Reported: 02/01/2012 Σ CGI ID No:5 Client:r Client Address: CLINICAL DATA: Lymphoma No CBC results provided. CLL Complete SM Report FINAL DIAGNOSIS: CD19+ B cell lymphoma, ZAP-70 + (44%), with borderline mutated IGHV, and gain of 3q. No TP53, ATM deletions. for t(11;14). COMMENTS: Correlation with comprehensive evaluation of clinical and pathological findings is recommended. FLOW CYTOMETRY: CD19+ B cell lymphoma, ZAP-70 + (44%). FISH: Normal CLL FISH Panel MOLECULAR: IGHV: Borderline mutated IGHV (close to 2%) MatBA: Positive for gain of 3q. Normal FISH: 11q13/14q32 Flow Plot ZAP-70 vs CD3 Date: 02/01/2012 Giumara, Ross J. Σ 12-000042 Page 1 of 1
Reported: 01/22/2012 Clinical Hx: Lymphoma/CLL CGI ID No: Client: Client Address: FLOW CYTOMETRY REPORT Interpretation: CD19+ B cell lymphoma, ZAP-70 + (44%). Comments: Correlation with comprehensive evaluation of clinical and pathological findings, as well as results of molecular and cytogenetic studies is recommended. Results: Viability (7-AAD): -% Differential Cell Count : Lymphocytes 90 % Monocytes 1 % Granulocytes 8 % Abnormal cell population: Present Abnormal cell size: Small (lymphocytes) Date: 01/22/2012 Giumara, Ross J. FC12-000045 Page 1 of 1
Subject ID: Sex: Male Specimen: PB Collected: Received: Reported: Female FLUORESCENCE IN SITU HYBRIDIZATION REPORT FISH Probes*: CLL [Abbott (Vysis), Inc.] PROBE SETS CHROMOSOME LOCI # CELLS ANALYZED ABNORMAL ABNORMAL CUTOFF VALUE (95% CI) RESULTS ISCN NOMENCLATURE 2009 11q22.3 (ATM), 17p13 (TP53) 300 --- Del 11q22.3 > 6.0% Del 17p13 > 7.0% No deletion ATM, TP53 nuc ish(atmx2,tp53x2) CEP 12/ 13q14(D13S319),13q34 200 --- Gain of 12 > 2.5% Del 13q > 5.5% Homozygous Del 13q > 1.5% Loss of 13 > 5.5% No deletion of 13q14/q34 No gain of CEP 12 detected nuc ish (D13S319x2,13q34x2,CEP12x2) 11q13 (CCND1), 14q32 (IGH) 200 --- t(11;14) > 1% No CCND1/IGH rearrangement nuc ish(ccnd1x2,ighx2) CEP6(D6Z1) 6q22-23(MYB) 200 --- Del(6q22-23) > 3% No deletion of MYB nuc ish(d6z1x2,mybx2) * FISH only testing is not equivalent to conventional cytogenetic analysis of the patient s specimen, and is limited to the specific probe(s) and their corresponding DNA locations (genes) only. FDA approved Normal/Abnormal determination cutoff. Summary: NORMAL CLL FISH PANEL RESULTS Final Interpretation: This FISH analysis showed normal signal patterns for 11q22.3/17p13, CEP12/13q14/13q34, 11q13/14q32, and CEP6/6q23 probes listed above in the CLL panel. Page 1 of 2
11q22.3/17p13.1 Cep12/13q14/13q34 11q13/14q32 CEP6/6q23 Representative images of the CGI CLL FISH panel showing nuclei with signal patterns for the 11q22 (2 green) /17p13 (2 red), CEP 12 (2 green)/ 13q14 (2 red)/13q34 (2 aqua), 11q13 (2 red)/14q32 (2 green), and CEP 6 (2 green)/6q23 (2 red) probe sets. FISH analysis can only identify abnormalities that are within the specific locus of the probe(s) used and may not detect small clonal populations of aberrant cells below the normal cut-off values; therefore, FISH results should be interpreted in the context of the patient s full clinical history and under most circumstances, in conjunction with metaphase chromosome analysis. D Pal Singh-Kahlon, PhD, FACMG, Director of Cytogenetics Date: 01/24/2012 FISH testing for HER2/neu (PathVysion), ALK for Non Small Cell Lung Cancer (NSCLC), MDS (for EGR1 (5q31)/5p15.2 probe), CLL (for TP53, ATM probes, D13S319 and D12Z3 probes), and X/Y probes for bone marrow (BM) transplant monitoring, have been approved by Food & Drug Administration (FDA) as in vitro diagnostic (IVD) tests. However, some other probes in use for MDS & CLL FISH tests are Analyte Specific Reagents (ASRs). In MDS, 7cen/7q31, cep8, 20q12, and MLL (11q23) probes are ASRs. In CLL c-myb(6q23), CCND1/IGH( t(11;14)) are ASRs. The tests utilizing analyte-specific reagents (ASRs) were developed & their performance characteristics determined by Cancer Genetics, Inc. as required by CLIA 88 regulations. They have not been cleared or approved by for specific uses by the Food and Drug Administration (FDA). The FDA has determined that such clearance or approval is not necessary. These tests are used for clinical purposes. Cancer Genetics Inc., 201, Route 17 North, Rutherford, NJ 070070. Phone Number : (888) 334-4988 CLIA#: 31D1038733 CAP LAP#: 7191582. Giumara, Ross J. FH12-000059 Page 2 of 2
Reported: 01/28/2012 Clinical Hx: Lymphoma Results: IGHV Family: V3-30 Functionality: CGI ID No: Client: Client Address: IGHV MUTATION ANALYSIS REPORT Mutation Frequency: 2.1% Productive Interpretation: Mutated (borderline mutated, see comment below) Comment: Patients with borderline IGHV mutation (close to 2% cut-off) have varying clinical outcome. Caution should be taken when interpreting clinical correlation. Description: The mutation status of the unique immunoglobulin gene (IGHV) rearrangement in the monoclonal proliferation of B-cells in chronic lymphocytic leukemia (CLL) is considered to have prognostic value. If mutations are detected at a level of 2% or higher in the sequenced V region of the clonal rearrangement, then the result is interpreted as Mutated. If mutations are detected at a level below 2% in the sequenced V region of the clonal rearrangement, then the result is interpreted as Unmutated. Those patients with a mutated IGHV gene usually have a less aggressive and more indolent disease, with longer overall survival. Those patients with an unmutated IGHV gene usually have a more aggressive disease and shorter overall survival. This assay utilizes PCR to detect a monoclonal IGHV rearrangement followed by sequence analysis to determine the specific IGHV family and mutation frequency. The sensitivity of the assay is 10%. Samples in which the monoclonal B-cells are present at less than 10%, a specific IGHV rearrangement will not be detected and will be reported as failures. Result Interpreted by: Weiyi Chen, Ph.D. Molecular Diagnostic Laboratory Director Date: 01/28/2012 Giumara, Ross J. M12-000085 Page 1 of 1
Reported: 02/01/2012 Clinical Hx: Lymphoma CGI ID No: Client: Client Address: MatBA -CLL/SLL ARRAY CGH REPORT Results: Genomic Aberration Loss of 8p Loss of 11q (ATM) Loss of 13q (MIR15A/16-1) Loss of 13q (RB1) Loss of 17p (TP53) Gain of 2p Gain of 3q Gain of 8q Gain of 12 Result for Aberration Positive Interpretation: Positive for the gain of 3q. Description: The gain and loss of specific genomic regions in the monoclonal proliferation of B-cells in chronic lymphocytic leukemia (CLL) are considered to have diagnostic and prognostic value. Loss of 13q at one locus (MIR15A/16-1) or both loci (MIR15A/16-1 and RB1) is observed in approximately 50% of CLL patients and as the sole abnormality is associated with a longer overall survival. Those patients with loss of 17p (TP53) or 11q (ATM) in general, have a shorter overall survival. Other aberrations are variously observed in CLL patients with suggested prognostic value. This assay utilizes microarray-based comparative genomic hybridization (array-cgh) to simultaneously detect the gain and loss of multiple loci in specimen DNA. Quantitative PCR is used to confirm the detected genomic gains and losses. The sensitivity of the assay is 30-40%. Samples in which the monoclonal B-cells are present at less than 30-40%, aberrations may not be detected and will be reported as no aberrations detected. Result Interpreted by: Weiyi Chen, Ph.D. Molecular Diagnostic Laboratory Director Date: 02/01/2012 Giumara, Ross J. M12-000084 Page 1 of 1