doi: /theoncologist

Size: px
Start display at page:

Download "doi: /theoncologist"

Transcription

1 Chromogenic In Situ Hybridization Histological and ThinPrep to Detect HER-2/neu Gene Amplification in -Processed Breast Cancer Fine-Needle Aspirates: A Sensitive and Practical Method in the Trastuzumab Era Amina Vocaturo, Flavia Novelli, Maria Benevolo, Giulia Piperno, Ferdinando Marandino, Anna Maria Cianciulli, Roberta Merola, Raffaele Perrone Donnorso, Isabella Sperduti, Simonetta Buglioni and Marcella Mottolese The Oncologist 2006, 11: doi: /theoncologist The online version of this article, along with updated information and services, is located on the World Wide Web at: Downloaded from by guest on April 28, 2014

2 Cancer Diagnostics and Molecular Pathology Chromogenic In Situ Hybridization to Detect HER-2/neu Gene Amplification in Histological and ThinPrep -Processed Breast Cancer Fine-Needle Aspirates: A Sensitive and Practical Method in the Trastuzumab Era Amina Vocaturo, a Flavia Novelli, a Maria Benevolo, a Giulia Piperno, a Ferdinando Marandino, a Anna Maria Cianciulli, b,d Roberta Merola, b Raffaele Perrone Donnorso, a Isabella Sperduti, c Simonetta Buglioni, a Marcella Mottolese a,d a Pathology Department, b Clinical Pathology, c Biostatistics Unit, d Breast Disease Management Team, Regina Elena Cancer Institute, Rome, Italy Key Words. Breast cancer HER-2 Chromogenic in situ hybridization Fluorescence in situ hybridization Liquid-based cytology Learning Objectives After completing this course, the reader will be able to: 1. Explain the importance of an accurate evaluation of HER-2 status to select breast cancer patients for trastuzumab therapy. 2. Describe the current methods that assess HER-2 status in breast cancer. 3. Discuss the advantages and limits of the CISH method for the detection of HER-2/neu gene amplification in histology and ThinPrep -processed breast cancer fine-needle aspirates. CME Access and take the CME test online and receive 1 AMA PRA Category 1 Credit at CME.TheOncologist.com Abstract The increasing evidence of trastuzumab efficacy in breast cancer (BC) patients means that an accurate and reproducible evaluation of HER-2 status is of paramount importance in histological and in cytological samples. Currently, the two main methods used to analyze HER- 2 amplification or overexpression are fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC). Although the two methods are strongly correlated for histological tissue, the evaluation of tumor morphology through FISH may be difficult and fluorescence fades quickly. These limitations can be overcome by chromogenic in situ hybridization (CISH), which can visualize the amplification product along with morphological features. In view of this, in the present study, we analyzed the usefulness of CISH on formalin-fixed, paraffin-embedded (FFPE) BC specimens and investigated whether CISH can be a valid technique in the determination of HER-2 status for fine-needle aspirates (FNAs) processed by liquid-based cytology. The results we obtained in a retrospective series of 111 FFPE BC specimens demonstrated good concordance between CISH and IHC and between CISH and FISH. The for- Correspondence: Amina Vocaturo, Ph.D., Regina Elena Cancer Institute, Via Elio Chianesi 53, Rome, Italy. Telephone: ; Fax: ; vocaturo@ifo.it Received May 8, 2006; accepted for publication July 20, AlphaMed Press /2006/$20.00/0 doi: /theoncologist The Oncologist 2006;11:

3 Vocaturo, Novelli, Benevolo et al. 879 mer concordance was comparable with that observed between FISH and IHC. When CISH was applied to a prospective series of 53 FNAs, from surgically removed BC, our data showed evidence of a higher concordance of results between liquid-based cytology and the companion FFPE tissues using CISH rather than HercepTest. Therefore, CISH analysis, which is a valuable and reproducible alternative to FISH for selecting breast cancer patients for trastuzumab therapy, can lower false-positive immunocytochemistry findings in ThinPrep -processed FNAs. The Oncologist 2006;11: Introduction The HER-2/neu oncogene is commonly overexpressed in several types of cancer. In breast cancer (BC) its overexpression, observed in 20% 30% of all cases, is closely related to gene amplification and associated with a worse prognosis [1]. Alteration of HER-2 status, known to be a predictive factor of the efficacy of specific chemotherapy [2] and hormonal therapy [3] regimens, has recently become of particular relevance in the current management of metastatic BC patients. This is a result of the availability of a recombinant, humanized monoclonal antibody directed against the extracellular domain of the HER-2 protein, trastuzumab (Herceptin ; Roche, Basel, Switzerland), which has significant antitumor activity both as a single agent and in combination with chemotherapy in advanced disease [4, 5]. More recently, it has been reported that, among patients with HER-2-positive BC, the addition of trastuzumab to chemotherapy in the neoadjuvant setting significantly increases the pathologic complete response rate [6] and, after adjuvant chemotherapy [7], strongly improves disease-free survival. Therefore, a laboratory assessment of HER-2 status, both on histological and cytological specimens, is becoming an increasingly important step in the optimal management of BC patients. Currently, two types of assays can be used for HER-2 evaluation: immunohistochemistry (IHC), which detects protein overexpression according to the U.S. Food and Drug Administration (FDA)-approved scoring staining system (0, 1+, 2+, 3+), and fluorescence in situ hybridization (FISH), which assesses gene amplification [8, 9]. Comparative studies of IHC and FISH have generally shown a high concordance rate between the two methods on primary or metastatic BC [8, 10]. Nevertheless, a high percentage of tumors displaying a 2+ score do not present concurrent gene amplification. In these discordant cases, HER-2/neu amplification appears to provide more prognostic information and better response to trastuzumab therapy than HER- 2 protein overexpression [11]. Indeed, although FISH is a very accurate and sensitive assay and is regarded as the gold standard test with a high sensitivity and specificity in detecting HER-2/neu amplification [12], this methodology has some disadvantages as it requires expensive microscope equipment, morphological features are difficult to visualize, and fluorescence fades quickly, so it does not provide a permanent record. Chromogenic in situ hybridization (CISH) is a recent methodology, which was introduced as an alternative to FISH. With this technique, HER-2/neu gene copies are detected using a permanent peroxidase reaction visualized by light microscopy, which makes it easy to observe both morphology and the amplification product [13, 14]. The present study analyzes the clinical usefulness of CISH, in our hospital-based practice, on a retrospective series of 111 histological samples of invasive BC. We compared the two in situ hybridization methods, CISH and FISH, with IHC by HercepTest (Dako, Milan, Italy) and CB11, which are the most frequently used reagents for detecting HER-2 overexpression. Moreover, in a prospective series of 53 BC patients, we investigated whether CISH can become a valid methodology for determining HER-2 status in fine-needle aspirates (FNAs) processed using ThinPrep (Cytyc, Rome, Italy) technology, comparing the results obtained with the corresponding formalin-fixed, paraffin-embedded (FFPE) BC specimens. Materials and Methods Patients A retrospective series of 111 primary infiltrating BC cases, diagnosed between 1992 and 2003, was selected from the Regina Elena Cancer Institute Pathology Department files. All these cases were diagnosed and treated at our institution and were referred for HER-2 testing between 2000 and We routinely performed an IHC study using a polyclonal antibody (pab) A0485 (HercepTest [Dako]). All tumors initially classified as scoring 2+ (36 cases) and 3+ (22 cases), according to the scoring guidelines of the FDA-approved HercepTest, were included in this study. Then, for statistical purposes, we randomly selected an overlapping number of negative cases (38 tumors with a score of 0 and 15 with a score of 1+) in order to obtain a similar number of positive and negative cases. A new hematoxylin and eosin stained section from each case was re-evaluated by two investigators (FM, GP) to assess whether the tissue specimens were still available for new tests. Then, all the 111 cases were processed for

4 880 CISH in Breast Cancer IHC with the monoclonal antibody (mab) CB11 (Novocastra, Menarini, Florence, Italy) for CISH and for FISH tests when not previously performed. A prospective series of 53 cytological samples was collected by performing FNA from surgically removed BC. Cytological specimens were processed using Thin- Prep methodology. At least five slides were obtained from each sample: one for Papanicolaou staining to verify the presence of neoplastic cells, one for immunocytochemical (ICC) staining (HercepTest), two for CISH, and one for FISH. Moreover, in order to confirm our cytological findings, the same procedures (i.e., IHC with HercepTest and CISH) were performed on FFPE BC tissues from the same 53 patients. The FISH analysis was performed on 26 of 53 matched cytological and histological samples. Cultured Cell Lines The SKBR3, BT474, and ZR751 cell lines were kindly provided by the Laboratory of Immunology, Regina Elena Cancer Institute, Rome, Italy. These BC-derived cell lines are reported to contain highly amplified (Fig. 1A), low amplified, and nonamplified HER-2/neu gene, respectively. The three cell lines were grown to confluence and cells were treated with trypsin-edta solution. After washing in phosphate buffer (0.1 M, ph 7.2), the cells were resuspended in CytoLyt (Cytyc), prepared by the ThinPrep technique, and then processed for ICC, CISH, and FISH, according to their respective technical procedures. These cells were then used as positive and negative controls. Tissue and FNA Preparation HER-2 overexpression and amplification were determined on 4-μm thick sections, which were cut from archival FFPE blocks and harvested on SuperFrost Plus slides (Menzel- Glaser, Braunschweig, Germany). FNA specimens were obtained from 53 surgically removed samples of BC within 30 minutes of resection, before tissue fixation. The aspirates were obtained using a 22-gauge needle and placed in CytoLyt for the fixation and preparation of ThinPrep slides according to the manufacturer s protocol. Corresponding BC tissue specimens were fixed in 10% neutral buffered formalin for hours according to routine procedures and embedded in paraffin. IHC/ICC on Histological and Cytological Specimens HER-2 overexpression was determined by the use of the pab A0485 and mab CB11. Both antibodies are directed against the intracytoplasmic domain of the HER-2 molecule. HercepTest had been previously performed according to the standard procedures described in the manufacturer s guide accompanying the kit. Immunoperoxidase staining for CB11 was carried out by pretreating the deparaffinized and rehydrated sections in a thermostatic bath at 96 C for 40 minutes in a 10 mm citrate buffer, ph 6. ThinPrep slides, after ethanol fixation and postfixation in buffered formalin, were washed in double-distilled water and processed as tissue specimens to determine HER-2 status. CB11 immunoreactivity was revealed using the HRP- LSAB2 system (Dako), employing 3-amino-9-ethylcarbazole (AEC, Dako) as chromogenic substrate. Sections were slightly counterstained with Mayer s hematoxylin and mounted in aqueous mounting medium (Glycergel ; Dako). For both specimen types, appropriate positive tissue and cell line controls were included in each run. Negative controls consisted of substituting the HER-2 primary antibody with normal rabbit serum (negative control reagent, Dako). Evaluation of the IHC/ICC results was done independently and blindly by two investigators (SB, MB). Figure 1. Chromogenic in situ hybridization in breast cancer cell line, histological and cytological samples. (A): HER- 2/neu gene amplification in ThinPrep -processed SKBR3 breast cancer cell line. (B): Invasive breast carcinoma showing chromosome 17 polysomy. (C F): Two ThinPrep -processed breast cancer fine-needle aspirates and the companion histological sections showing no HER-2/neu amplification (two signals) (C, D) and HER-2/neu amplification (multiple signals) (E, F) in tumor cell nuclei. Scale bar = 30 μm. The Oncologist

5 Vocaturo, Novelli, Benevolo et al. 881 HER-2 overexpression was determined as defined in the HercepTest kit guide: scores of 0 or 1+ were considered negative, a score of 2+ was considered weak positive, and a score of 3+ was considered strong positive. To qualify for 2+ scoring, complete membrane staining on more than 10% of tumor cells at a weak intensity had to be observed. In the majority of 3+ cases, at least 80% of the tumor cells demonstrated intense and homogeneous cell membrane staining. FISH Procedure on Histological and Cytological Specimens FISH was performed using the PathVysion assay kit (Vysis, Inc., Downers Grove, IL), which includes two directly labeled DNA probes: a locus-specific probe for the HER-2/neu gene labeled with SpectrumOrange (LSI HER-2) and an alpha satellite probe that targets the centromere region of chromosome 17 labeled with SpectrumGreen chromosome enumeration probe (CEP 17). The assay was performed according to the manufacturer s instructions. In brief, after deparaffinization, FFPE sections were incubated in the pretreatment solution (80 C, 10 minutes) and then digested with protease (37 C, 15 minutes). FNA slides, obtained by the ThinPrep technique, were immersed in Carnoy s fixative and stored at 20 C before FISH analysis. Cytological specimens were dehydrated before applying LSI HER-2/CEP 17 probe and then were hybridized overnight at 37 C. After washing, slides were counterstained with 4ʹ,6-diamidino-2-phenylindole (DAPI) and analyzed under a fluorescence microscope. An average of 200 nuclei was enumerated in the invasive component of tumor tissue and an average of tumor cells was enumerated in cytological preparations. HER-2/ neu gene copy number, chromosome 17 copy number, and the average HER-2/neu gene to chromosome 17 signal ratio were reported as FISH genetic variables. Samples with a ratio value 2.0 were considered to be amplified. CISH Procedure on Histological and Cytological Specimens FFPE sections were deparaffinized, dehydrated, air dried, and heated in boiling Tissue Heat Pretreatment Buffer for 15 minutes using a SPoT-Light FFPE reagent kit (Zymed, Histoline, Milan, Italy). The enzymatic digestion was performed using SPoT-Light FFPE digestion enzyme (Zymed) for 10 minutes at room temperature (RT). FNA ThinPrep slides, prefixed in 96% ethanol, were postfixed in 99% ethanol, air dried, and then immersed in 2X standard saline preheated citrate solution (SSC; Zymed) at 37 C for 1 hour. After dehydration, histological and cytological slides were air dried and the ready-to-use double-stranded DNA digoxygenin-labeled HER2 probe (Zymed) or the biotinlabeled Chromosome 17 Centromeric Probe (Zymed) were applied. The denaturation was performed by incubating the slides, covered with a CISH coverslip (Zymed), on a 96 C heating block for 5 minutes, and the hybridization was performed by placing the slides in a humidity chamber at 37 C overnight. After removing the coverslips, a stringent wash was performed in 0.5X SSC at 80 C for 5 minutes. The endogenous peroxidase activity and unspecific staining were blocked by applying 3% H 2 O 2 and the CAS-Block (Zymed), respectively. A mouse antidigoxygenin antibody was added to slides hybridized with HER-2 probe for 45 minutes at RT followed by incubation with a polymerized peroxidase-goat anti-mouse antibody (Dako) for 45 minutes at RT. On FFPE tissue slides, the colorimetric signal of the Chromosome 17 Centromeric Probe was improved by incubating the slides with a mouse antibiotin antibody (Dako) for 45 minutes at 37 C. For cytological slides, the Chromosome 17 Centromeric Probe was detected employing a horseradish peroxidase (HRP)-streptavidin for 45 minutes at RT. A 3,3ʹ-diaminobenzidine (DAB) chromogen substrate system was used to generate a sensitive signal that could be viewed with a transmission light (brightfield) microscope after hematoxylin counterstaining. Evaluation of CISH Results Amplification was considered to be high when more than 10 copies, or large clusters, of the HER-2/neu gene were present in >50% of cancer cell nuclei, whereas 6 10 copies of the HER-2/neu gene, or a small HER-2/neu gene cluster, in the same percentage of cells were considered to be low amplification. Tumors were not considered to be amplified when 1 5 copies of the HER-2/neu gene were identified per nucleus. In the group of tumors with low amplification (6 10 copies/nucleus), an adjacent serial section was hybridized with the Chromosome 17 Centromeric Probe to detect polysomic cases (Fig. 1B). CISH signals were read by two investigators (MM, AV) independently of the results of the other two assays. Statistics Correlation among tests (IHC, ICC, CISH, and FISH) as well as interobserver agreement were estimated using the kappa test. For the IHC comparison of HER-2 overexpression, categories 0 and 1+ were grouped together as negative results and categories 2+ and 3+ were considered positive results. p <.05 was considered statistically significant. Specificity, sensitivity, negative and positive predictive value (NPV and PPV), concordance, and the 95% confidence interval (CI) of the CISH assay were estimated considering FISH as the gold standard.

6 882 CISH in Breast Cancer Results IHC Analysis, CISH, and FISH on Histological Specimens When negative (0/1+) and positive (2+/3+) scores were grouped together, a good agreement between CB11 and HercepTest immunoreactivity (κ = 0.80; 95% CI, 69% 92%; p <.0001) was evidenced in our retrospective series of 111 BC specimens. In particular, 52 cases were negative and 49 were positive with both reagents, whereas 10 cases were discordant. As summarized in Table 1, on the basis of this good agreement, we correlated HER-2 protein overexpression detected by HercepTest with gene amplification detected by the two genetic assays. The concordance between HER-2 protein overexpression and HER-2/neu gene amplification, evaluated either by FISH or CISH, was 100% in the 53 cases with a 0/1+ score. In the 36 BC cases displaying a 2+ score, 17 (47%) were not amplified and 19 (53%) were amplified, with a complete concordance between FISH and CISH. However, of the 22 score 3+ tumors, 20 (91%) were identified as amplified by FISH and 14 were identified as amplified by CISH (64%). As summarized in Table 2, when FISH was chosen as the gold standard, the overall concordance between FISH and CISH was 95%. We observed 72 nonamplified and 33 amplified BC cases with both genetic assays, with six false-negative and no false-positive cases, with a κ coefficient for the interassay agreement of 0.88 (95% CI, 78% 97%; p <.0001). Sensitivity for CISH was 85% (95% CI, 73% 95%), specificity was 100% (95% CI, 100% 100%), PPV was 100% (95% CI, 100% 100%), and NPV was 92% (95% CI, 86% 98%). All six FISH CISH discordant tumors demonstrated a 3+ score when the HercepTest was used, whereas using the mab CB11, three cases displayed a 3+ and three displayed a 2+ score (data not shown). In order to analyze the concordance among the three tests used in this study (namely, IHC, FISH, and CISH) in more detail, we statistically evaluated the overall concordance between tumors presenting either 0/1+ or 3+ scores and HER-2/neu gene amplification status assessed both by CISH and FISH. Score 2+ tumors, in fact, known to be amplified only in a limited number of cases, were excluded from this statistical analysis. As shown in Figure 2, comparing protein status by HercepTest with gene amplification by CISH and FISH, we evidenced 89% (κ = 0.71; p <.0001; 95% CI, 82% 96%) and 97% (κ = 0.93, p <.0001; CI, 94% 100%) concordance, respectively. On the other hand, the concordance between CB11 immunoreactivity and gene amplification was 90% for CISH (κ = 0.74; p <.0001, 95% CI, 83% 96%) and 93% for FISH (κ = 0.84; p <.0001; 95% CI, 88% 98%). ICC Analysis, CISH, and FISH on Cytological Specimens Once the CISH procedure on FFPE tissues was validated, we analyzed whether this method could also be successfully applied to cytological specimens. To this aim, a consecutive series of 53 FNAs sampled from histologically assessed BC was processed by ThinPrep methodology as described in Materials and Methods. Before performing Table 1. Correlation between HER-2 overexpression and HER-2/neu gene amplification by FISH and CISH in histological tissues FISH CISH HercepTest score No amplification (%) Amplification (%) No amplification (%) Amplification (%) Total 0 38 (100) 0 (0) 38 (100) 0 (0) (100) 0 (0) 15 (100) 0 (0) (47) 19 (53) 17 (47) 19 (53) (9) 20 (91) 8 (36) 14 (64) 22 Total Abbreviations: CISH, chromogenic in situ hybridization; FISH, fluorescence in situ hybridization. Table 2. Comparison between HER-2/neu gene amplification detected by FISH and CISH in histological tissues FISH CISH No amplification (%) Amplification (%) Total No amplification 72 (65) 6 (5) 78 Amplification 0 33 (30) 33 Total Concordance, 95%; κ = 0.88; p < ; sensitivity, 85%; specificity, 100%; positive predictive value, 100%; negative predictive value, 92%. Abbreviations: CISH, chromogenic in situ hybridization; FISH, fluorescence in situ hybridization. The Oncologist

7 Vocaturo, Novelli, Benevolo et al. 883 CISH, cytological specimens and the corresponding FFPE BC tissues were tested for HER-2 protein overexpression using the HercepTest. The ICC staining, performed in the 53 prospective FNAs, evidenced 37 (70%) cases that were HER-2 negative (score 0 or 1+) and 16 (30%) that were positive. The 37 HER-2-negative cytological cases were also negative in histological sections and none of them presented HER-2/ neu gene amplification. When we focused on the 16 HER- 2-positive FNAs (Table 3), we observed that four of these cases (three score 3+ and one score 2+) were negative in histological sections (score 1+). These discordant cytological cases were interpreted as being ICC false positive because they were nonamplified with the CISH and FISH tests, both in FNA and in FFPE samples. Six of the remaining 12 cases presented overlapping results using ICC, IHC, CISH, and FISH in both cytology and histology (all BC cases were not Figure 2. Comparison among immunohistochemistry, chromogenic in situ hybridization (CISH), and fluorescence in situ hybridization (FISH) on breast cancer histological samples. Percentage concordance between negative (0/1+) and immunohistochemically strongly positive (3+) tumors and HER- 2/neu gene amplification evaluated by CISH and FISH. (A): HercepTest versus CISH (concordance, 89%). (B): HercepTest versus FISH (concordance, 97%). (C): CB11 versus CISH (concordance, 90%). (D): CB11 versus FISH (concordance, 93%). amplified, including one case scored 3+ by ICC and IHC). Another three cases, ICC and IHC positive (two score 3+, one score 2+), were amplified by CISH both in liquid-based cytology (LBC) FNAs and in histological tissues. In the latter tumors, because of the paucity of cellular material, we were unable to determine the HER-2/neu gene amplification status by FISH in FNAs and we did not perform FISH on the companion FFPE specimens. The last three BC specimens were all amplified both by CISH and FISH. In particular, using FISH, we found a HER-2/neu to CEP 17 ratios of 3, 2.5, and 4 in cytological preparations and of 5.1, 2.8, and 3.6 in the corresponding histological specimens. A lower HER- 2 score was detected with ICC (score 2+) than with IHC (score 3+). Therefore, of 53 FNAs, 46 showed overlapping ICC results with those obtained on histological tissues (concordance, 87%; κ = 0.69; p <.0001; 95% CI, 78% 96%). Concerning CISH, a 100% concordance was observed between LBC and histological samples. In fact, 45 of the 53 BC specimens (85%) did not show HER-2/neu gene amplification (1 4 gene copies/nucleus; Fig. 1C, D), and eight (15%) presented low (6 10 gene copies/nucleus) or high (>10 gene copies/nucleus) amplification both in FNAs and in the corresponding FFPE tissues (Fig. 1E, F). In order to further validate the CISH test in LBC, 26 of the 53 FNAs containing an adequate number of neoplastic cells (>50) and the companion histological sections were also tested by FISH. The two series showed completely overlapping results, with 3 of 26 cases (12%) amplified and 23 cases (88%) nonamplified with the two genetic tests (CISH and FISH), both on the cytological and the corresponding histological samples with 100% of concordance. (Data not shown.) Interobserver Agreement Interobserver agreement, calculated by κ value, indicated an excellent concordance for both IHC (κ = 0.96; 95% CI, Table 3. Analysis of HER-2/neu gene amplification by CISH and FISH in the 16 HercepTest-positive breast cancer fine-needle aspirates: comparison with the corresponding histological tissues LBC FNA Histological tissues No. of cases ICC score CISH FISH IHC score CISH FISH 3 3 No A No A 1 No A No A 1 2 No A No A 1 No A No A 1 3 No A No A 3 No A No A 5 2 No A No A 2 No A No A 2 3 A NV 3 A ND 1 2 A NV 2 A ND 3 2 A A 3 A A Abbreviations: A, amplification; CISH, chromogenic in situ hybridization; FISH, fluorescence in situ hybridization; ICC, immunocytochemistry; IHC, immunohistochemistry; LBC FNA, liquid-based cytology fine-needle aspirates; No A, no amplification; ND, not done; NV, not valuable.

8 884 CISH in Breast Cancer 94.7% 100%) and CISH (κ = 0.94; 95% CI, 94.3% 100%), p < Those cases in which there was a discrepancy between the two readers were resolved by reviewing the cases using a multiheaded microscope and reaching a consensus. Discussion Overexpression and/or amplification of the HER-2/neu gene, as stated by the last St. Gallen expert consensus meeting [15], represents a new risk category in BC, fundamental in the algorithm for the selection of adjuvant systemic treatments. Moreover, the widely documented link between HER-2 status and response to endocrine [3] and anthracycline-based [2] therapies, as well as the recent trials for trastuzumab efficacy in the adjuvant [7] and neoadjuvant [6] settings, make mandatory a standardized, accurate, and reproducible determination of HER-2 status on both histological and cytological specimens. Although FISH is in general considered the gold standard methodology for detecting HER-2/neu gene amplification, the recent development of the CISH assay provides an attractive alternative to FISH, especially in laboratories that are familiar with peroxidase-based staining. In recent years, the advantages of CISH, which allows a simultaneous evaluation of gene copy number, tumor cells, and detailed surrounding tissue morphology on the same histological slide, have been widely reported by several authors [14, 16, 17]. In contrast, data on the comparative use of ICC and CISH on cytological specimens and the corresponding FFPE tissues are minimal [18, 19]. To address this issue, in our study, we first validated CISH on a retrospective series of FFPE BC samples, subsequently focusing on the feasibility of CISH in a prospective series of BC FNAs processed using ThinPrep technology. In FFPE samples, complete agreement between FISH and CISH results was demonstrated for IHC scores 0, 1+, and 2+. A lower concordance for score 3+ tumors was found because six tumors were amplified by FISH and nonamplified by CISH. A careful re-examination of the six CISH FISH discordant tumors revealed that all these cases were collected from 1992 to 1998, and we can hypothesize that these FFPE specimens were ineffective as a result of preanalytic problems, such as bad tissue preservation or a poorly standardized formalin fixation method. The latter could represent a limit of this new assay when compared with FISH [12, 20]. The current CISH procedure is based on a single color detection of one probe and does not allow correction for HER-2/neu amplification caused by chromosome 17 polysomy, as in the dual-color FISH test provided by Vysis. Nevertheless, in our series, we did not find any CISH false-positive cases. This is probably because of the fact that, in tumors displaying 6 10 HER-2/neu gene copies/nucleus, an adjacent consecutive section was consistently hybridized with CEP 17 for comparison. Although Sartelet et al. [19] reported that polysomy is a rare event in cases with a low gene copy number, we strongly feel that this is a crucial point, because, without adjusting CISH results for chromosome 17 copy number, a higher number of discordant interpretations could result in borderline cases [14, 17, 20 22]. In agreement with the majority of studies reporting a concordance between FISH and CISH of 83% 100% [12 14, 16, 17, 20, 23, 24], in our series of FFPE tissues, the interassay concordance between the two genetic tests was 95% (κ = 0.88). This accurate histological validation of the CISH procedure provided the technical basis to test the feasibility of CISH in LBC, comparing our results on histologically matched tumors. We had two main aims. First, we assessed the potential value of CISH in cytological samples. Second, we verified whether LBC, which offers the possibility of adjunctive investigations on the same homogeneous samples, may be a suitable methodology to detect HER-2/neu gene amplification by CISH as already demonstrated by FISH [10]. Although FNA procedures are ever more frequently used instead of tissue biopsy to obtain diagnostic cellular material in primary inoperable or metastatic BC, to the best of our knowledge, only two studies have analyzed the CISH assay on conventional smears [18] or ThinPrep cytology [19] in parallel with the companion FFPE tissues. Moreover, neither of the two studies compared CISH with FISH findings. In summary, our results demonstrate that CISH is easily applicable to LBC with a good correlation to results obtained with the corresponding histological sections, and a complete agreement with FISH. This issue is of major importance, because CISH could be particularly helpful in overcoming the problem of false-positive findings with ICC, one of the major criticisms of this methodology [25, 26]. In our series, we found four positive cases (three score 3+, one score 2+) using ICC with LBC that were negative using IHC (score 1+) on the companion FFPE tissues and nonamplified by CISH both in ThinPrep and histological samples. These findings seem to indicate that CISH may lower the ICC false positivity that is probably a result of the poorly assessed scoring system in cytological material. FNA BC cells, in fact, lack an architectural tissue structure, which makes it difficult, in most cases, to apply the FDA-approved histological scoring system to cytopathology. These interpretative difficulties may lead to a great variability in evaluating the immunocolorimetric signals, which consequently become highly subjective. In contrast, The Oncologist

9 Vocaturo, Novelli, Benevolo et al. 885 CISH allows a more objective interpretation of HER-2/neu gene status, mainly with LBC. This technical procedure, in fact, different from conventional smears, permits a monolayer cell assessment. Consequently, tumor cells appear cleaner and chromogenic dots are easier to score because of intact nuclei. Considering the fact that in all ancillary tests the technical thoroughness is very important, in our cytological series, preanalytical procedures were followed very carefully, allowing a complete agreement of results between CISH and FISH (100% concordance). Conclusion In conclusion, our results provide evidence that CISH analysis performed on FFPE tissues should be considered a viable alternative to FISH analysis for selecting IHC 2+ scored patients for trastuzumab therapy. Moreover, in cytological specimens, it provides stronger and more consistent correlation than protein evaluation with the corresponding FFPE tissue samples. Based on the present results, which need to References 1 Ross JS, Fletcher JA, Linette GP et al. The Her-2/neu gene and protein in breast cancer 2003: biomarker and target of therapy. The Oncologist 2003;8: Piccart M, Lohrisch C, Di Leo A et al. The predictive value of HER2 in breast cancer. Oncology 2001;61(suppl 2): De Laurentiis M, Arpino G, Massarelli E et al. A meta-analysis on the interaction between HER-2 expression and response to endocrine treatment in advanced breast cancer. Clin Cancer Res 2005;11: Slamon DJ, Leyland-Jones B, Shak S et al. Use of chemotherapy plus a monoclonal antibody against HER2 for metastatic breast cancer that overexpress HER2. N Engl J Med 2001;344: Vogel CL, Cobleigh MA, Tripathy D et al. Efficacy and safety of trastuzumab as a single agent in first-line treatment of HER2-overexpressing metastatic breast cancer. J Clin Oncol 2002;20: Buzdar AU, Ibrahim NK, Francis D et al. Significantly higher pathologic complete remission rate after neoadjuvant therapy with trastuzumab, paclitaxel, and epirubicin chemotherapy: results of a randomized trial in human epidermal growth factor receptor 2-positive operable breast cancer. J Clin Oncol 2005;23: Piccart-Gebhart MJ, Procter M, Leyland-Jones B et al. Trastuzumab after adjuvant chemotherapy in HER2-positive breast cancer. N Engl J Med 2005;353: Jimenez RE, Wallis T, Tabasczka P et al. Determination of Her-2/Neu status in breast carcinoma: comparative analysis of immunohistochemistry and fluorescent in situ hybridization. Mod Pathol 2000;13: Birner P, Oberhuber G, Stani J et al. Evaluation of the United States Food and Drug Administration-approved scoring and test system of HER-2 protein expression in breast cancer. Clin Cancer Res 2001;7: Beatty BG, Bryant R, Wang W et al. HER2/neu detection in fine-needle aspirates of breast cancer: fluorescence in situ hybridization and immunocytochemical analysis. Am J Clin Pathol 2004;122: be confirmed by a larger study, we suggest that CISH can be considered a useful, simple, and reproducible method that is less expensive and appears to be a valuable alternative to FISH. Moreover, CISH can avoid false-positive ICC findings, increasing the accuracy of determining HER-2 status in breast cancer cytological samples processed using Thin- Prep methodology. Acknowledgments We thank Dr. Pier Giorgio Natali for his continuous support and helpful discussion, Michael Kenyon for the formal revision of the manuscript, and Maria Assunta Fonsi for secretarial assistance. This study has been supported by Ministero della Salute, Lega Italiana per la Lotta contro i Tumori, AIRC, on behalf of the Breast Disease Management Team (DMT). Disclosure of Potential Conflicts of Interest The authors indicate no potential conflicts of interest. 11 Pauletti G, Dandekar S, Rong H et al. Assessment of methods for tissuebased detection of the HER-2/neu alteration in human breast cancer: a direct comparison of fluorescence in situ hybridization and immunohistochemistry. J Clin Oncol 2000;18: Gupta D, Middleton LP, Whitaker MJ et al. Comparison of fluorescence and chromogenic in situ hybridization for detection of HER-2/neu oncogene in breast cancer. Am J Clin Pathol 2003;119: Tanner M, Gancberg D, Di Leo A et al. Chromogenic in situ hybridization: a practical alternative for fluorescence in situ hybridization to detect HER-2/neu oncogene amplification in archival breast cancer samples. Am J Pathol 2000;157: Isola J, Tanner M, Forsyth A et al. Interlaboratory comparison of HER-2 oncogene amplification as detected by chromogenic and fluorescence in situ hybridization. Clin Cancer Res 2004;10: Goldhirsch A, Glick JH, Gelber RD et al. Meeting highlights: international expert consensus on the primary therapy of early breast cancer Ann Oncol 2005;16: Arnould L, Denoux Y, MacGrogan G et al. Agreement between chromogenic in situ hybridisation (CISH) and FISH in the determination of HER2 status in breast cancer. Br J Cancer 2003;88: Bhargava R, Lal P, Chen B. Chromogenic in situ hybridization for the detection of HER-2/neu gene amplification in breast cancer with an emphasis on tumors with borderline and low-level amplification: does it measure up to fluorescence in situ hybridization? Am J Clin Pathol 2005;123: Kim GY, Oh YL. Chromogenic in situ hybridization analysis of HER- 2/neu status in cytological samples of breast carcinoma. Cytopathology 2004;15: Sartelet H, Lagonotte E, Lorenzato M et al. Comparison of liquid based cytology and histology for the evaluation of HER-2 status using immunostaining and CISH in breast carcinoma J Clin Pathol 2005;58: Zhao J, Wu R, Au A et al. Determination of HER2 gene amplification by chromogenic in situ hybridization (CISH) in archival breast carcinoma. Mod Pathol 2002;15:

10 886 CISH in Breast Cancer 21 Wang S, Hossein Saboorian M, Frenkel EP et al. Aneusomy 17 in breast cancer: its role in HER-2/neu protein expression and implication for clinical assessment of HER-2/neu status. Mod Pathol 2002;15: Gong Y, Gilcrease M, Sneige N. Reliability of chromogenic in situ hybridization for detecting HER-2 gene status in breast cancer: comparison with fluorescence in situ hybridization and assessment of interobserver reproducibility. Mod Pathol 2005;18: Dandachi N, Dietze O, Hauser-Kronberger C. Chromogenic in situ hybridization: a novel approach to a practical and sensitive method for the detection of HER2 oncogene in archival human breast carcinoma. Lab Invest 2002;82: Loring P, Cummins R, O Grady A et al. HER2 positivity in breast carcinoma: a comparison of chromogenic in situ hybridization with fluorescence in situ hybridization in tissue microarrays, with targeted evaluation of intratumoral heterogeneity by in situ hybridization. Appl Immunohistochem Mol Morphol 2005;13: Nizzoli R, Bozzetti C, Crafa P et al. Immunocytochemical evaluation of HER-2/neu on fine-needle aspirates from primary breast carcinomas. Diagn Cytopathol 2003;28: Bedard YC, Pollett AF, Leung SW et al. Assessment of thin-layer breast aspirates for immunocytochemical evaluation of HER2 status. Acta Cytol 2003;47: The Oncologist

11 Citations This article has been cited by 7 HighWire-hosted articles:

Priti Lal, MD, 1 Paulo A. Salazar, 1 Clifford A. Hudis, MD, 2 Marc Ladanyi, MD, 1 and Beiyun Chen, MD, PhD 1. Abstract

Priti Lal, MD, 1 Paulo A. Salazar, 1 Clifford A. Hudis, MD, 2 Marc Ladanyi, MD, 1 and Beiyun Chen, MD, PhD 1. Abstract Anatomic Pathology / DUAL- VS SINGLE-COLOR SCORING IN IMMUNOHISTOCHEMICAL AND FISH HER-2 TESTING HER-2 Testing in Breast Cancer Using Immunohistochemical Analysis and Fluorescence In Situ Hybridization

More information

Department of Pathology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

Department of Pathology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA & 2005 USCAP, Inc All rights reserved 0893-3952/05 $30.00 www.modernpathology.org Reliability of chromogenic in situ hybridization for detecting HER-2 gene status in breast cancer: comparison with fluorescence

More information

Comparison of Immunohistochemical and Fluorescence In Situ Hybridization Assessment of HER-2 Status in Routine Practice

Comparison of Immunohistochemical and Fluorescence In Situ Hybridization Assessment of HER-2 Status in Routine Practice Anatomic Pathology / ASSESSMENT OF HER-2 STATUS Comparison of Immunohistochemical and Fluorescence In Situ Hybridization Assessment of HER-2 Status in Routine Practice Michelle Dolan, MD, 1 and Dale Snover,

More information

HER-2/neu amplification detected by fluorescence in situ hybridization in fine needle aspirates from primary breast cancer

HER-2/neu amplification detected by fluorescence in situ hybridization in fine needle aspirates from primary breast cancer Original article Annals of Oncology 13: 1398 1403, 2002 DOI: 10.1093/annonc/mdf217 HER-2/neu amplification detected by fluorescence in situ hybridization in fine needle aspirates from primary breast cancer

More information

Journal of Breast Cancer

Journal of Breast Cancer ORIGINAL ARTICLE Journal of Breast Cancer J Breast Cancer 2009 December; 12(4): 235-40 DOI: 10.4048/jbc.2009.12.4.235 Comparison of Silver-Enhanced in situ Hybridization and Fluorescence in situ Hybridization

More information

HER2 CISH pharmdx TM Kit Interpretation Guide Breast Cancer

HER2 CISH pharmdx TM Kit Interpretation Guide Breast Cancer P A T H O L O G Y HER2 CISH pharmdx TM Kit Interpretation Guide Breast Cancer FROM CERTAINTY COMES TRUST For in vitro diagnostic use HER2 CISH pharmdx Kit HER2 CISH pharmdx Kit is intended for dual-color

More information

Comparison of Fluorescence and Chromogenic In Situ Hybridization for Detection of HER-2/neu Oncogene in Breast Cancer

Comparison of Fluorescence and Chromogenic In Situ Hybridization for Detection of HER-2/neu Oncogene in Breast Cancer Anatomic Pathology / HER-2 DETECTION BY CISH IN BREAST CANCER Comparison of Fluorescence and Chromogenic In Situ Hybridization for Detection of HER-2/neu Oncogene in Breast Cancer Deepali Gupta, MD, 1

More information

T he HER2/neu type 1 tyrosine kinase growth factor

T he HER2/neu type 1 tyrosine kinase growth factor 710 ORIGINAL ARTICLE HER2 amplification status in breast cancer: a comparison between immunohistochemical staining and fluorescence in situ hybridisation using manual and automated quantitative image analysis

More information

Considerable advances in the therapy of breast cancer

Considerable advances in the therapy of breast cancer HER-2/neu Status in Breast Cancer Metastases to the Central Nervous System Kelly C. Lear-Kaul, MD; Hye-Ryoung Yoon, MD; Bette K. Kleinschmidt-DeMasters, MD; Loris McGavran, PhD; Meenakshi Singh, MD Context.

More information

KEY WORDS: Breast carcinoma, c-erbb2, Fluorescent. Mod Pathol 2001;14(11):

KEY WORDS: Breast carcinoma, c-erbb2, Fluorescent. Mod Pathol 2001;14(11): HER-2/neu in Breast Cancer: Interobserver Variability and Performance of Immunohistochemistry with 4 Antibodies Compared with Fluorescent In Situ Hybridization Thomas A. Thomson, M.D., Malcolm M. Hayes,

More information

HER2 FISH pharmdx TM Interpretation Guide - Breast Cancer

HER2 FISH pharmdx TM Interpretation Guide - Breast Cancer P A T H O L O G Y HER2 FISH pharmdx TM Interpretation Guide - Breast Cancer For In Vitro Diagnostic Use FDA approved as an aid in the assessment of patients for whom Herceptin TM (trastuzumab) treatment

More information

Comparison of in situ hybridization methods for the assessment of HER-2/neu gene amplification status in breast cancer using a tissue microarray

Comparison of in situ hybridization methods for the assessment of HER-2/neu gene amplification status in breast cancer using a tissue microarray reports of practical oncology and radiotherapy 1 7 ( 2 0 1 2 ) 44 49 Available online at www.sciencedirect.com jo u r n al hom epage: http://www.elsevier.com/locate/rpor Original article Comparison of

More information

Dr. dr. Primariadewi R, SpPA(K)

Dr. dr. Primariadewi R, SpPA(K) Curriculum Vitae Dr. dr. Primariadewi R, SpPA(K) Education : Medical Doctor from UKRIDA Doctoral Degree from Faculty of Medicine University of Indonesia Pathologist Specialist and Consultant from Faculty

More information

Immunohistochemical (IHC) HER-2/neu and Fluorescent- In Situ Hybridization (FISH) Gene Amplification of Breast Cancer in Indian Women

Immunohistochemical (IHC) HER-2/neu and Fluorescent- In Situ Hybridization (FISH) Gene Amplification of Breast Cancer in Indian Women Comparison of IHC and FISH for HER-2/neu Status of Breast Cancer in Indian Women RESEARCH COMMUNICATION Immunohistochemical (IHC) HER-2/neu and Fluorescent- In Situ Hybridization (FISH) Gene Amplification

More information

Reviewer's report. Version: 1 Date: 24 May Reviewer: Cathy Moelans. Reviewer's report:

Reviewer's report. Version: 1 Date: 24 May Reviewer: Cathy Moelans. Reviewer's report: Reviewer's report Title: Validation of HER2 testing with core needle biopsy specimens from primary breast cancers in terms of interobserver reproducibility and concordance with surgically resected specimens

More information

Comparison on Cell Block, Needle-Core, and Tissue Block Preparations

Comparison on Cell Block, Needle-Core, and Tissue Block Preparations Immunohistochemical Detection of Estrogen Receptor, Progesterone Receptor, and Human Epidermal Growth Factor Receptor 2 Expression in Breast Carcinomas Comparison on Cell Block, Needle-Core, and Tissue

More information

IT S ABOUT TIME. IQFISH pharmdx Interpretation Guide THREEHOURSTHIRTYMINUTES. HER2 IQFISH pharmdxtm. TOP2A IQFISH pharmdxtm

IT S ABOUT TIME. IQFISH pharmdx Interpretation Guide THREEHOURSTHIRTYMINUTES. HER2 IQFISH pharmdxtm. TOP2A IQFISH pharmdxtm I N T E R P R E TAT I O N IQFISH pharmdx Interpretation Guide TM HER2 IQFISH pharmdxtm TOP2A IQFISH pharmdxtm Breast carcinoma (FFPE) stained with HER2 IQFISH pharmdx Breast carcinoma (FFPE) stained with

More information

Welcome! HER2 TESTING DIAGNOSTIC ACCURACY 4/11/2016

Welcome! HER2 TESTING DIAGNOSTIC ACCURACY 4/11/2016 HER2 TESTING DIAGNOSTIC ACCURACY Can t We Finally Get It Right? Allen M. Gown, M.D. Medical Director and Chief Pathologist PhenoPath Laboratories Seattle, Washington Clinical Professor of Pathology University

More information

B reast carcinomas are the most frequent tumours in

B reast carcinomas are the most frequent tumours in 864 ORIGINAL ARTICLE Comparison of liquid based cytology and histology for the evaluation of HER-2 status using immunostaining and CISH in breast carcinoma H Sartelet, E Lagonotte, M Lorenzato, I Duval,

More information

Three Hours Thirty Minutes

Three Hours Thirty Minutes INTERPRETATION HER2 IQFISH pharmdx TM Interpretation Guide Three Hours Thirty Minutes it s about time Breast carcinoma (FFPE) stained with HER2 IQFISH pharmdx Gastric cancer (FFPE) stained with HER2 IQFISH

More information

Comparative Analysis of Methods Used in Breast Cancer HER2 and Sentinel Lymph Node Diagnosis

Comparative Analysis of Methods Used in Breast Cancer HER2 and Sentinel Lymph Node Diagnosis SHORT THESIS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY (Ph.D.) Comparative Analysis of Methods Used in Breast Cancer HER2 and Sentinel Lymph Node Diagnosis by Monika Francz MD Supervisor: Zoltán Szöllősi

More information

Technical Advance. Chromogenic in Situ Hybridization

Technical Advance. Chromogenic in Situ Hybridization American Journal of Pathology, Vol. 157, No. 5, November 2000 Copyright American Society for Investigative Pathology Technical Advance Chromogenic in Situ Hybridization A Practical Alternative for Fluorescence

More information

Instant Quality FISH. The name says it all.

Instant Quality FISH. The name says it all. PRODUCT INFORMATION HER2 IQFISH pharmdx Instant Quality FISH Instant Quality FISH. The name says it all. HER2 IQFISH pharmdx IQ: Instant Quality every time. HER2 IQFISH pharmdx stains of a HER2 non-amplified

More information

CANCER. Clinical Validation of Breast Cancer Predictive Markers

CANCER. Clinical Validation of Breast Cancer Predictive Markers Clinical Validation of Breast Cancer Predictive Markers David Hicks, MD Loralee McMahon, MS, HTL(ASCP) CANCER The human body is composed of billions of highly regulated cells Cancer cells no longer respond

More information

HER2 Gene Protein Assay Is Useful to Determine HER2 Status and Evaluate HER2 Heterogeneity in HER2 Equivocal Breast Cancer

HER2 Gene Protein Assay Is Useful to Determine HER2 Status and Evaluate HER2 Heterogeneity in HER2 Equivocal Breast Cancer HER2 Gene Protein Assay Is Useful to Determine HER2 Status and Evaluate HER2 Heterogeneity in HER2 Equivocal Breast Cancer Yanjun Hou, MD, PhD, 1 Hiroaki Nitta, PhD, 2 and Zaibo Li, MD, PhD 1 From the

More information

HER2+ Breast Cancer Review of Biologic Relevance and Optimal Use of Diagnostic Tools

HER2+ Breast Cancer Review of Biologic Relevance and Optimal Use of Diagnostic Tools Anatomic Pathology / HER2: BIOLOGIC RELEVANCE AND DIAGNOSIS HER2+ Breast Cancer Review of Biologic Relevance and Optimal Use of Diagnostic Tools David G. Hicks, MD, 1 and Swati Kulkarni, MD 2 Key Words:

More information

Dako IT S ABOUT TIME. Interpretation Guide. Agilent Pathology Solutions. ALK, ROS1 and RET IQFISH probes (Dako Omnis) MET IQFISH probe (Dako Omnis)

Dako IT S ABOUT TIME. Interpretation Guide. Agilent Pathology Solutions. ALK, ROS1 and RET IQFISH probes (Dako Omnis) MET IQFISH probe (Dako Omnis) INTERPRETATION Dako Agilent Pathology Solutions IQFISH Interpretation Guide ALK, ROS1 and RET IQFISH probes (Dako Omnis) MET IQFISH probe (Dako Omnis) IT S ABOUT TIME For In Vitro Diagnostic Use ALK, ROS1,

More information

# Best Practices for IHC Detection and Interpretation of ER, PR, and HER2 Protein Overexpression in Breast Cancer

# Best Practices for IHC Detection and Interpretation of ER, PR, and HER2 Protein Overexpression in Breast Cancer #1034 - Best Practices for IHC Detection and Interpretation of ER, PR, and HER2 Protein Overexpression in Breast Cancer Richard W. Cartun, MS, PhD Andrew Ricci, Jr, MD Department of Pathology Hartford

More information

HER2 status assessment in breast cancer. Marc van de Vijver Academic Medical Centre (AMC), Amsterdam

HER2 status assessment in breast cancer. Marc van de Vijver Academic Medical Centre (AMC), Amsterdam HER2 status assessment in breast cancer Marc van de Vijver Academic Medical Centre (AMC), Amsterdam 13e Bossche Mamma Congres 17 th June 2015 Modern cancer therapies are based on sophisticated molecular

More information

MEDICAL POLICY. Proprietary Information of YourCare Health Plan

MEDICAL POLICY. Proprietary Information of YourCare Health Plan MEDICAL POLICY SUBJECT: HER-2 TESTING IN INVASIVE BREAST OR PAGE: 1 OF: 7 If the member's subscriber contract excludes coverage for a specific service it is not covered under that contract. In such cases,

More information

HER2/neu Amplification in Breast Cancer Stratification by Tumor Type and Grade

HER2/neu Amplification in Breast Cancer Stratification by Tumor Type and Grade Anatomic Pathology / HER2/NEU AMPLIFICATION IN BREAST CANCER HER2/neu Amplification in Breast Cancer Stratification by Tumor Type and Grade Elise R. Hoff, MD, Raymond R. Tubbs, DO, Jonathan L. Myles, MD,

More information

CME/SAM. Abstract. Anatomic Pathology / HER2/neu Results in Breast Cancer

CME/SAM. Abstract. Anatomic Pathology / HER2/neu Results in Breast Cancer Anatomic Pathology / HER2/neu Results in Breast Cancer Effect of Ischemic Time, Fixation Time, and Fixative Type on HER2/neu Immunohistochemical and Fluorescence In Situ Hybridization Results in Breast

More information

Product Introduction

Product Introduction Product Introduction Product Codes: HCL026, HCL027 and HCL028 Contents Introduction to HER2 2 HER2 immunohistochemistry 3 Cell lines as controls 5 HER2 Analyte Control DR IHC 7 HER2 Analyte Control DR

More information

Prediction of HER2 gene status in Her2 2 þ invasive breast cancer: a study of 108 cases comparing ASCO/CAP and FDA recommendations

Prediction of HER2 gene status in Her2 2 þ invasive breast cancer: a study of 108 cases comparing ASCO/CAP and FDA recommendations & 2009 USCAP, Inc All rights reserved 0893-3952/09 $32.00 www.modernpathology.org Prediction of HER2 gene status in Her2 2 þ invasive breast cancer: a study of 108 cases comparing ASCO/CAP and FDA recommendations

More information

Assessment Run B HER2 IHC

Assessment Run B HER2 IHC Assessment Run B24 2017 HER2 IHC Material The slide to be stained for HER2 comprised the following 5 materials: IHC: HER2 Score* (0, 1+, 2+, 3+) FISH: HER2 gene/chr 17 ratio** 1. Breast carcinoma, no.

More information

HER2 ISH (BRISH or FISH)

HER2 ISH (BRISH or FISH) Assessment Run H14 2018 HER2 ISH (BRISH or FISH) Material Table 1. Content of the multi-block used for the NordiQC HER2 ISH assessment, run H14 HER2 IHC* IHC score Dual - SISH** FISH*** FISH*** HER2/chr17

More information

Journal of Breast Cancer

Journal of Breast Cancer Journal of Breast Cancer ORIGINAL ARTICLE J Breast Cancer 2011 December; 14(4): 276-282 Silver-Enhanced In Situ Hybridization as an Alternative to Fluorescence In Situ Hybridization for Assaying HER2 Amplification

More information

Approximately one third of all cancer-related deaths in

Approximately one third of all cancer-related deaths in ORIGINAL ARTICLE Comparison Between Epidermal Growth Factor Receptor (EGFR) Gene Expression in Primary Non-small Cell Lung Cancer (NSCLC) and in Fine-Needle Aspirates from Distant Metastatic Sites Cecilia

More information

S Wang, M H Saboorian, E Frenkel, L Hynan, S T Gokaslan, R Ashfaq

S Wang, M H Saboorian, E Frenkel, L Hynan, S T Gokaslan, R Ashfaq 374 Department of Pathology, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75235-9072, USA S Wang M H Saboorian R Ashfaq Department of Internal Medicine, University

More information

On May 4 and 5, 2002, the College of American Pathologists

On May 4 and 5, 2002, the College of American Pathologists College of American Pathologists Conference Conference Summary, Strategic Science Symposium Her-2/neu Testing of Breast Cancer Patients in Clinical Practice Richard J. Zarbo, MD, DMD; M. Elizabeth H. Hammond,

More information

Enhanced Accuracy and Reliability of HER-2/neu Immunohistochemical Scoring Using Digital Microscopy

Enhanced Accuracy and Reliability of HER-2/neu Immunohistochemical Scoring Using Digital Microscopy Anatomic Pathology / HER-2 IMMUNOHISTOCHEMICAL SCORING RELIABILITY Enhanced Accuracy and Reliability of HER-2/neu Immunohistochemical Scoring Using Digital Microscopy Kenneth Bloom, MD, 1 and Douglas Harrington,

More information

HER2 status in breast cancer: experience of a Spanish National Reference Centre

HER2 status in breast cancer: experience of a Spanish National Reference Centre Clin Transl Oncol (2011) 13:000-000 DOI RESEARCH ARTICLES HER2 status in breast cancer: experience of a Spanish National Reference Centre Marta Cuadros Carlos Cano Francisco Javier López Paloma Talavera

More information

Nitta et al. Diagnostic Pathology 2012, 7:60

Nitta et al. Diagnostic Pathology 2012, 7:60 Nitta et al. Diagnostic Pathology 2012, 7:60 METHODOLOGY Open Access A gene-protein assay for human epidermal growth factor receptor 2 (HER2): brightfield tricolor visualization of HER2 protein, the HER2

More information

Determination of HER2 Amplification by In Situ Hybridization. When Should Chromosome 17 Also Be Determined?

Determination of HER2 Amplification by In Situ Hybridization. When Should Chromosome 17 Also Be Determined? Anatomic Pathology / FISH f o r HER2: Wh e n to Use Ch r o m o s o m e 17 Determination of HER2 Amplification by In Situ Hybridization When Should Chromosome 17 Also Be Determined? John M.S. Bartlett,

More information

Instant Quality FISH. The name says it all.

Instant Quality FISH. The name says it all. COMPANION DIAGNOSTICS Instant Quality FISH Instant Quality FISH. The name says it all. IQ: Instant Quality every time. Breast carcinoma stained with : Triple filter showing Blue DAPI colors nuclei, FITC

More information

HER2/neu Evaluation of Breast Cancer in 2019

HER2/neu Evaluation of Breast Cancer in 2019 HER2/neu Evaluation of Breast Cancer in 2019 A.A. Sahin, M.D. Professor of Pathology and Translation Molecular Pathology Section Chief of Breast Pathology ERBB2 (HER2) Background 185-kDa membrane protein

More information

Kristen E. Muller, DO, Jonathan D. Marotti, MD, Vincent A. Memoli, MD, Wendy A. Wells, MD, and Laura J. Tafe, MD

Kristen E. Muller, DO, Jonathan D. Marotti, MD, Vincent A. Memoli, MD, Wendy A. Wells, MD, and Laura J. Tafe, MD AJCP / Original Article Impact of the 2013 ASCO/CAP HER2 Guideline Updates at an Academic Medical Center That Performs Primary HER2 FISH Testing Increase in Equivocal Results and Utility of Reflex Immunohistochemistry

More information

Available online

Available online Available online http://breast-cancer-research.com/content/9/5/r64 Vol 9 No 5 Research article Comparison of different commercial kits for HER2 testing in breast cancer: looking for the accurate cutoff

More information

MEDICAL POLICY. Proprietary Information of Excellus Health Plan, Inc. A nonprofit independent licensee of the BlueCross BlueShield Association

MEDICAL POLICY. Proprietary Information of Excellus Health Plan, Inc. A nonprofit independent licensee of the BlueCross BlueShield Association MEDICAL POLICY SUBJECT: HER-2 TESTING IN INVASIVE BREAST OR PAGE: 1 OF: 7 If a product excludes coverage for a service, it is not covered, and medical policy criteria do not apply. If a commercial product,

More information

Oncologist. The. Breast Cancer

Oncologist. The. Breast Cancer The Oncologist Breast Cancer Comparison of HER-2 and Hormone Receptor Expression in Primary Breast Cancers and Asynchronous Paired Metastases: Impact on Patient Management VALENTINA GUARNERI, a SIMONA

More information

Brief Formalin Fixation and Rapid Tissue Processing Do Not Affect the Sensitivity of ER Immunohistochemistry of Breast Core Biopsies

Brief Formalin Fixation and Rapid Tissue Processing Do Not Affect the Sensitivity of ER Immunohistochemistry of Breast Core Biopsies Brief Formalin Fixation and Rapid Tissue Processing Do Not Affect the Sensitivity of ER Immunohistochemistry of Breast Core Biopsies Victoria Sujoy, MD, Mehrdad Nadji, MD, and Azorides R. Morales, MD From

More information

Immunohistochemistry in Breast Pathology- Brief Overview of the Technique and Applications in Breast Pathology

Immunohistochemistry in Breast Pathology- Brief Overview of the Technique and Applications in Breast Pathology SMGr up Immunohistochemistry in Breast Pathology- Brief Overview of the Technique and Applications in Breast Pathology Bhanumathi K Rao 1 * 1 Department of Biochemistry, JSS Medical College, a constituent

More information

Contributions to Anatomic Pathology, over the years

Contributions to Anatomic Pathology, over the years Contributions to Anatomic Pathology, over the years Anatomic Pathology, part 1 G.B. Morgagni Xavier Bichat Rudolf Wirchow Anatomic Pathology, part 1 Anatomic pathology materials: morphological samples

More information

Genetic heterogeneity in HER2/neu testing by fluorescence in situ hybridization: a study of 2522 cases

Genetic heterogeneity in HER2/neu testing by fluorescence in situ hybridization: a study of 2522 cases Modern Pathology () 5, 683 688 & USCAP, Inc. All rights reserved 893-395/ $3. 683 Genetic heterogeneity in HER/neu testing by fluorescence in situ hybridization: a study of 5 cases Martin C Chang,,3, Janet

More information

Assessment Run B HER-2

Assessment Run B HER-2 Assessment Run B1 2006 HER-2 The slide to be stained for HER-2 comprised: 1. Cell line JIMT-1 (Amplified)* 2. Cell line MDA-453 (Amplified) 3. Cell line MCF-7 (Not amplified) 4. Cell line BT474 (Amplified)

More information

Oncology Genetics: Cytogenetics and FISH 17/09/2014

Oncology Genetics: Cytogenetics and FISH 17/09/2014 Oncology Genetics: Cytogenetics and FISH 17/09/2014 Chris Wragg Head of Oncology Genomics, BGL BGL Bristol Genetics Laboratory (BGL) CPA accredited Genetics laboratory serving a core population of 4-5million

More information

Evaluation of c-erbb-2 overexpression and Her-2/neu gene copy number heterogeneity in Barrett s adenocarcinoma

Evaluation of c-erbb-2 overexpression and Her-2/neu gene copy number heterogeneity in Barrett s adenocarcinoma 25 Evaluation of c-erbb-2 overexpression and Her-2/neu gene copy number heterogeneity in Barrett s adenocarcinoma Axel Walch a,, Karin Bink b, Peter Gais a, Stefan Stangl a, Peter Hutzler a, Michaela Aubele

More information

University of Groningen

University of Groningen University of Groningen Validation of the 4B5 rabbit monoclonal antibody in determining Her2/neu status in breast cancer van der Vegt, Bert; de Bock, Gertruida H; Bart, Jos; Zwartjes, N.G.; Wesseling,

More information

Assessment of Her-2/neu Overexpression in Primary Breast Cancers and Their Metastatic Lesions: An Immunohistochemical Study

Assessment of Her-2/neu Overexpression in Primary Breast Cancers and Their Metastatic Lesions: An Immunohistochemical Study Annals o f Clinical & Laboratory Science, vol. 30, no. 3, 2000 259 Assessment of Her-2/neu Overexpression in Primary Breast Cancers and Their Metastatic Lesions: An Immunohistochemical Study Shahla Masood

More information

Impact of Polysomy 17 on HER-2/neu Immunohistochemistry in Breast Carcinomas without HER-2/neu Gene Amplification

Impact of Polysomy 17 on HER-2/neu Immunohistochemistry in Breast Carcinomas without HER-2/neu Gene Amplification Journal of Molecular Diagnostics, Vol. 5, No. 3, August 2003 Copyright American Society for Investigative Pathology and the Association for Molecular Pathology Impact of Polysomy 17 on HER-2/neu Immunohistochemistry

More information

Received 04 November 2008; Accepted in revision 09 January 2009; Available online 20 January 2009

Received 04 November 2008; Accepted in revision 09 January 2009; Available online 20 January 2009 Int J Clin Exp Pathol (2009) 2, 476-480 www.ijcep.com/ijcep811001 Original Article Immunohistochemical Detection of Estrogen and Progesterone Receptor and HER2 Expression in Breast Carcinomas: Comparison

More information

NIH Public Access Author Manuscript Cancer Epidemiol Biomarkers Prev. Author manuscript; available in PMC 2011 January 1.

NIH Public Access Author Manuscript Cancer Epidemiol Biomarkers Prev. Author manuscript; available in PMC 2011 January 1. NIH Public Access Author Manuscript Published in final edited form as: Cancer Epidemiol Biomarkers Prev. 2010 January ; 19(1): 144 147. doi:10.1158/1055-9965.epi-09-0807. Feasibility Study for Collection

More information

Pepsin Solution ready-to-use

Pepsin Solution ready-to-use SIE HABEN DIE VISION, WIR HABEN DIE SUBSTANZ. Pepsin Solution Single component Pepsin Solution: only one component refrigerator stable Pepsin is a commonly used digestive enzyme for immunohistochemical

More information

What kind of material should we use for ICC in our daily routine. Torill Sauer Department of Pathology, Akershus University Hospital

What kind of material should we use for ICC in our daily routine. Torill Sauer Department of Pathology, Akershus University Hospital What kind of material should we use for ICC in our daily routine Torill Sauer Department of Pathology, Akershus University Hospital Diversity of preparing cytological material Cell block Direct smears

More information

American Society of Cytopathology Core Curriculum in Molecular Biology

American Society of Cytopathology Core Curriculum in Molecular Biology American Society of Cytopathology Core Curriculum in Molecular Biology American Society of Cytopathology Core Curriculum in Molecular Biology Chapter 6 Fluorescence in situ Hybridization (FISH) Principles

More information

Assessment Run B HER-2 IHC. HER-2/chr17 ratio**

Assessment Run B HER-2 IHC. HER-2/chr17 ratio** Assessment Run B2 20 HER-2 IHC Material The slide to be stained for HER-2 comprised the following 5 tissues: IHC HER-2 Score* (0, +, 2+,3+) FISH HER-2/chr7 ratio**. Breast ductal carcinoma 0..3 2. Breast

More information

Characterization and significance of MUC1 and c-myc expression in elderly patients with papillary thyroid carcinoma

Characterization and significance of MUC1 and c-myc expression in elderly patients with papillary thyroid carcinoma Characterization and significance of MUC1 and c-myc expression in elderly patients with papillary thyroid carcinoma Y.-J. Hu 1, X.-Y. Luo 2, Y. Yang 3, C.-Y. Chen 1, Z.-Y. Zhang 4 and X. Guo 1 1 Department

More information

Immunohistochemical Determination of HER-2/neu Expression in Invasive Breast Carcinoma

Immunohistochemical Determination of HER-2/neu Expression in Invasive Breast Carcinoma Anatomic Pathology / HER-2/NEU IN BREAST CARCINOMA Immunohistochemical Determination of HER-2/neu Expression in Invasive Breast Carcinoma Russell Vang, MD, 1 Linda D. Cooley, MD, 1 Wilbur R. Harrison,

More information

Assessment Run B HER2 IHC

Assessment Run B HER2 IHC Assessment Run B26 208 HER2 IHC Material The slide to be stained for HER2 comprised the following 5 materials: IHC: HER2 Score* (0, +, 2+, 3+) FISH: HER2 gene/chr 7 ratio**. Breast carcinoma, no. 2+..3

More information

Interpretation Manual - Gastric or Gastroesophageal Junction Adenocarcinoma. PD-L1 IHC 22C3 pharmdx is FDA-approved for in vitro diagnostic use

Interpretation Manual - Gastric or Gastroesophageal Junction Adenocarcinoma. PD-L1 IHC 22C3 pharmdx is FDA-approved for in vitro diagnostic use Interpretation Manual - Gastric or Gastroesophageal Junction Adenocarcinoma PD-L1 IHC 22C3 pharmdx is FDA-approved for in vitro diagnostic use For countries outside of the United States, see the local

More information

Assessment of Breast Cancer with Borderline HER2 Status Using MIP Microarray

Assessment of Breast Cancer with Borderline HER2 Status Using MIP Microarray Assessment of Breast Cancer with Borderline HER2 Status Using MIP Microarray Hui Chen, Aysegul A Sahin, Xinyan Lu, Lei Huo, Rajesh R Singh, Ronald Abraham, Shumaila Virani, Bal Mukund Mishra, Russell Broaddus,

More information

HercepTest for the Dako Autostainer Code K5207

HercepTest for the Dako Autostainer Code K5207 HercepTest for the Dako Autostainer Code K5207 9th edition For immunocytochemical staining. The kit is for 50 tests (100 slides). (126659-002) P04088US_02_K520721-5/2016.05 p. 1/54 Contents Page Intended

More information

Quality assurance and quality control in pathology in breast disease centers

Quality assurance and quality control in pathology in breast disease centers Quality assurance and quality control in pathology in breast disease centers Judith Sandbank M.D. Pathology Assaf-Harofeh Medical Center ISRAEL jsandbank@asaf.health.gov.il 1 st IBDC, 28 th January, 2011

More information

Template for Reporting Results of Biomarker Testing of Specimens From Patients With Carcinoma of the Breast

Template for Reporting Results of Biomarker Testing of Specimens From Patients With Carcinoma of the Breast Template for Reporting Results of Biomarker Testing of Specimens From Patients With Carcinoma of the Breast Version: Template Posting Date: January 2018 Includes requirements from the 2017 CAP Accreditation

More information

Guideline. Associated Documents ASCO CAP 2018 GUIDELINES and SUPPLEMENTS -

Guideline. Associated Documents ASCO CAP 2018 GUIDELINES and SUPPLEMENTS - Guideline Subject: ASCO CAP 2018 HER2 Testing for Breast Cancer Guidelines - Recommendations for Practice in Australasia Approval Date: December 2018 Review Date: December 2022 Review By: HER2 testing

More information

(A) PCR primers (arrows) designed to distinguish wild type (P1+P2), targeted (P1+P2) and excised (P1+P3)14-

(A) PCR primers (arrows) designed to distinguish wild type (P1+P2), targeted (P1+P2) and excised (P1+P3)14- 1 Supplemental Figure Legends Figure S1. Mammary tumors of ErbB2 KI mice with 14-3-3σ ablation have elevated ErbB2 transcript levels and cell proliferation (A) PCR primers (arrows) designed to distinguish

More information

HercepTest TM Code K5204

HercepTest TM Code K5204 HercepTest TM Code K5204 9th edition For immunocytochemical staining. The kit is for 35 tests (70 slides). (126559-002) P04086US_02_K520421-5/US/2016.05 p. 1/55 Contents Intended Use... 4 Summary and Explanation

More information

Evolution of Pathology

Evolution of Pathology 1 Traditional pathology Molecular pathology 2 Evolution of Pathology Gross Pathology Cellular Pathology Morphologic Pathology Molecular/Predictive Pathology Antonio Benivieni (1443-1502): First autopsy

More information

Journal of the Egyptian Nat. Cancer Inst., Vol. 22, No. 4, December: , 2010

Journal of the Egyptian Nat. Cancer Inst., Vol. 22, No. 4, December: , 2010 Journal of the Egyptian Nat. Cancer Inst., Vol. 22, No. 4, December: 27-225, 2 Assessment of the Reliability of Immunocytochemical Detection of Estrogen and Progesterone Receptors Status on the Cytological

More information

Immunohistochemistry on Fluid Specimens: Technical Considerations

Immunohistochemistry on Fluid Specimens: Technical Considerations Immunohistochemistry on Fluid Specimens: Technical Considerations Blake Gilks Dept of Pathology University of British Columbia, Vancouver, BC, Canada Disclosures None Learning Objectives At the end of

More information

Breast Cancer Interpretation Guide

Breast Cancer Interpretation Guide Breast Cancer Interpretation Guide UCT D O R P NEW ERBB2/ C E P S ht e ZytoLig lor Prob o C l a u 2D D17S12 ng to the i d r o c c a ting for re-tes idelines 2013 ASCO Gu Breast Cancer Interpretation Guide

More information

In some regions of the world, breast cancer mortality beginning to fall due to earlier

In some regions of the world, breast cancer mortality beginning to fall due to earlier Original rticle Keykhosro Mardanpour (MD) 1 Mahtab Rahbar (MD) *2 Sedigheh Khazaei (MSc) 3 Mohsen ghaitasvand (MD) 3 1- Department of Orthopedics, Imam Reza Hospital, Kermanshah University of Medical Sciences,

More information

ACCURACY OF IMMUNOHISTOCHEMISTRY IN EVALUATION

ACCURACY OF IMMUNOHISTOCHEMISTRY IN EVALUATION POL J PATHOL 2011; 2: 95-100 ACCURACY OF IMMUNOHISTOCHEMISTRY IN EVALUATION OF MALIGNANT PLEURAL AND PERITONEAL EFFUSIONS FERESHTEH ENSANI, FARNAZ NEMATIZADEH, GITI IRVANLOU Department of Cytology, Cancer

More information

What is HER2 positive breast cancer in 2018? Updated ASCO-CAP guidelines. Giuseppe Viale University of Milan European Institute of Oncology

What is HER2 positive breast cancer in 2018? Updated ASCO-CAP guidelines. Giuseppe Viale University of Milan European Institute of Oncology What is HER2 positive breast cancer in 2018? Updated ASCO-CAP guidelines Giuseppe Viale University of Milan European Institute of Oncology Mission accomplished! First alarming results Breast Intergroup

More information

Controversies in HER2 Oncogene Testing: What Constitutes a True Positive Result in Patients With Breast Cancer?

Controversies in HER2 Oncogene Testing: What Constitutes a True Positive Result in Patients With Breast Cancer? Controversies in HER2 Oncogene Testing: What Constitutes a True Positive Result in Patients With Breast Cancer? Michael F. Press, MD, PhD; Yanling Ma, MD; Susan Groshen, PhD; Guido Sauter, MD, PhD; and

More information

erb-b2 Amplification by Fluorescence In Situ Hybridization in Breast Cancer Specimens Read as 2+ in Immunohistochemical Analysis

erb-b2 Amplification by Fluorescence In Situ Hybridization in Breast Cancer Specimens Read as 2+ in Immunohistochemical Analysis Anatomic Pathology / ERB-B2 FISH+ AND IMMUNOHISTOCHEMICALLY 2+ erb-b2 Amplification by Fluorescence In Situ Hybridization in Breast Cancer Specimens Read as 2+ in Immunohistochemical Analysis Chieh Lan,

More information

Results you can trust

Results you can trust PRODUCT I NF OR MAT ION pharmdx Results you can trust The first and only FDA-approved PD-L1 test to assess the magnitude of treatment effect on progression-free survival in melanoma patients from OPDIVO

More information

Preanalytic Variables in Cytology: Lessons Learned from Next Generation Sequencing

Preanalytic Variables in Cytology: Lessons Learned from Next Generation Sequencing @Sinchita_Roy #USCAP2017 #PulmPath #IAmUSCAP #insitupathologists Preanalytic Variables in Cytology: Lessons Learned from Next Generation Sequencing Sinchita Roy-Chowdhuri, MD, PhD Department of Pathology

More information

Version 2 of these Guidelines were drafted in response to published updated ASCO/CAP HER2 test Guideline Recommendations-

Version 2 of these Guidelines were drafted in response to published updated ASCO/CAP HER2 test Guideline Recommendations- Introduction: These guidelines represent systematically developed statements to assist in the provision of quality assured HER2 testing in breast and gastric/ gastro-oesophageal carcinoma. They are based

More information

T here are convincing data to support the hypothesis that a

T here are convincing data to support the hypothesis that a 367 ORIGINAL ARTICLE Status of the p53, p16, RB1, and HER-2 genes and chromosomes 3, 7, 9, and 17 in advanced bladder cancer: correlation with adjacent mucosa and pathological parameters M Gallucci, F

More information

CME/SAM. Abstract. Anatomic Pathology / Image Analysis of HER2 Immunostaining

CME/SAM. Abstract. Anatomic Pathology / Image Analysis of HER2 Immunostaining Anatomic Pathology / Image Analysis of HER2 Immunostaining Image Analysis of HER2 Immunohistochemical Staining Reproducibility and Concordance With Fluorescence In Situ Hybridization of a Laboratory-Validated

More information

Potential Value of Hormone Receptor Assay in Carcinoma In Situ of Breast

Potential Value of Hormone Receptor Assay in Carcinoma In Situ of Breast Potential Value of Hormone Receptor Assay in Carcinoma In Situ of Breast ROBERT BARNES, M.D. AND SHAHLA MASOOD, M.D. The estrogen receptor (ER) expression of invasive breast cancer has been extensively

More information

CME/SAM ABSTRACT. AJCP / Original Article

CME/SAM ABSTRACT. AJCP / Original Article Clinicopathologic Significance of the Intratumoral Heterogeneity of HER2 Gene Amplification in HER2- Positive Breast Cancer Patients Treated With Adjuvant Trastuzumab Hee Jin Lee, MD, PhD, 1 Joo Young

More information

A Study to Evaluate the Effect of Neoadjuvant Chemotherapy on Hormonal and Her-2 Receptor Status in Carcinoma Breast

A Study to Evaluate the Effect of Neoadjuvant Chemotherapy on Hormonal and Her-2 Receptor Status in Carcinoma Breast Original Research Article A Study to Evaluate the Effect of Neoadjuvant Chemotherapy on Hormonal and Her-2 Receptor Status in Carcinoma Breast E. Rajesh Goud 1, M. Muralidhar 2*, M. Srinivasulu 3 1Senior

More information

Study of Melanin Bleaching After Immunohistochemistry of Melanin-containing Tissues. Hongwu Shen, MD and Wenqiao Wu, MD

Study of Melanin Bleaching After Immunohistochemistry of Melanin-containing Tissues. Hongwu Shen, MD and Wenqiao Wu, MD TECHNICAL ARTICLE Study of Melanin Bleaching After Immunohistochemistry of Melanin-containing Tissues Hongwu Shen, MD and Wenqiao Wu, MD Abstract: Melanin may interfere with immunohistochemical staining.

More information

HRP cytochemistry. Division of Radiooncology, Deutsches Krebsforschungszentrum, Heidelberg, Germany

HRP cytochemistry. Division of Radiooncology, Deutsches Krebsforschungszentrum, Heidelberg, Germany HRP cytochemistry WOLF D. KUHLMANN, M.D. Division of Radiooncology, Deutsches Krebsforschungszentrum, 69120 Heidelberg, Germany A range of substrates is available for the cytochemical staining of peroxidase

More information

Next-Generation Immunohistochemistry: Multiplex tissue imaging with mass cytometry

Next-Generation Immunohistochemistry: Multiplex tissue imaging with mass cytometry Nat Met, April 2014 Nat Med, April 2014 Next-Generation Immunohistochemistry: Multiplex tissue imaging with mass cytometry Journal Club Timo Böge Overview Introduction Conventional Immunohistochemistry

More information

Detection of Anaplastic Lymphoma Kinase (ALK) gene in Non-Small Cell lung Cancer (NSCLC) By CISH Technique

Detection of Anaplastic Lymphoma Kinase (ALK) gene in Non-Small Cell lung Cancer (NSCLC) By CISH Technique Cancer and Clinical Oncology; Vol. 7, No. 1; 2018 ISSN 1927-4858 E-ISSN 1927-4866 Published by Canadian Center of Science and Education Detection of Anaplastic Lymphoma Kinase (ALK) gene in Non-Small Cell

More information