Genotyping of Human Papillomavirus

Size: px

Start display at page:

Download "Genotyping of Human Papillomavirus"

Deirdre Lloyd
6 years ago
Views:

1 VisinArray Arrays fr DNA analysis Gentyping f Human Papillmavirus mplete C r u Y frm n i t u l S ctin a r t x E DNA sis! y l a n A t Simultaneus Detectin f 41 HPV Types High Sensitivity Easy t Handle

Over the last years the relevance f HPV in the histry f rpharyngeal cancers has becme mre and mre imprtant which is indicated by a dramatically risen number f cancers f the ral cavity and pharynx

2 Imprtance f HPV Testing The human papillmavirus (HPV) has been cnclusively identified as the majr risk factr fr cervical cancer. It is the third mst cmmn cancer in wmen wrldwide, with an estimated number f 530,000 new cases and 280,000 deaths each year. Over the last years the relevance f HPV in the histry f rpharyngeal cancers has becme mre and mre imprtant which is indicated by a dramatically risen number f cancers f the ral cavity and pharynx linked t HPV. Cervical Cancer Prgressin Initial HPV infectin requires virin integratin int the basal epithelial cells. Whereas mst HPV infectins are transient and are cleared by the immune system, infectin with HPV high risk types may lead t cancer when infectin persists. Expsure Transient Infectin Persistant Infectin Infectin Prgressin Invasin Nrmal Cervix Clearance Prductive Infectin Regressin Precursr Lesin Cancer Cytlgy? Cytlgy? HPV Gentype? Cytlgy? HPV Gentype? Cytlgy? HPV Gentype? Duratin mnths 24 mnths 7-15 years VisinArray HPV Chip Fast and Reliable Gentyping The VisinArray HPV Chip 1.0 is intended t be used with the VisinArray Analysis Package fr the qualitative detectin and gentyping f PCR amplificates f 41 clinically relevant HPV gentypes High Risk HPV Types detected by the VisinArray HPV Chip a 68b IS39 MM Prbably High Risk HPV Types detected by the VisinArray HPV Chip a 68a b MM4 82 IS39 68b (IS39) 82 (MM4) Lw Risk HPV Types detected by the VisinArray HPV Chip High Risk Prbably High Risk Lw Risk Guide Dts/Psitive Cntrl (CP8304) 83 (MM7) 84 (MM8) References: Bumba LM, et al. (2014) BMC Public Health 14: 1320 Clmb N, et al. (2012) Ann Oncl 23 Suppl 7: vii27-32 Crw JM, et al. (2012) Nature 488: S2-S3 IARC (2012) Bilgical Agents. IARC Mngr Eval Carcing Risks Hum, 100B: Pljak M, et al. (2016) J Clin Virl 76 Suppl 1: S3-S13

1. Sample Cllectin Fr the detectin f HPV gentypes with the

fr DNA extractin: Frmalin-fixed, paraffin-embedded (FFPE)

Amplificatin and Labeling in a PCR Bitinylated primers are

the bitinylated sequences hybridize t cmplementary DNA

Step 3: Detectin and Visualizatin Specifically bund and

3 1. Sample Cllectin Fr the detectin f HPV gentypes with the VisinArray HPV system the fllwing raw material can be used fr DNA extractin: Frmalin-fixed, paraffin-embedded (FFPE) tissue samples Cervical swab/brush specimen Liquid based cytlgy specimen (ThinPrep ) 2. Detectin & Differentiatin f 41 HPV Types Step 1: Amplificatin and Labeling in a PCR Bitinylated primers are used t amplify and label different sectins f the L1 regin f the HPV genme. The human HLA-DQA1 gene is als amplified and serves as a PCR psitive cntrl and as a genmic cntrl. Step 2: Hybridizatin n the Glass Chip After amplificatin, the bitinylated sequences hybridize t cmplementary DNA capture sequences n the glass chip. Step 3: Detectin and Visualizatin Specifically bund and bitinylated sequences are visualized by secndary marking with a streptavidinperxidase cnjugate and a staining with tetramethylbenzidine. After clr develpment, evaluatin is perfrmed using the VisinArray Analyzer Sftware.

4 3. Wrkflw Schedule This is a cndensed prtcl fr the VisinArray methd and shuld nt replace the instructin fr use! 0 Min. PCR Fr the PCR the VisinArray Primer Kit is used The master mix is prepared by using the dntp/dutp Slutin, HPV Primer Mix, VisinArray PreCise Taq Plymerase, VisinArray Uracil-DNA Glycsylase and the crrespnding reactin buffers Alternatively: The VisinArray HPV PreCise Master Mix can be used DNA sample is added t the master mix Hybridizatin PCR prduct and Hybridizatin Slutin are mixed well Mix is applied nt the VisinArray Chip 30 Min. Duratin: 30 min Stringency Wash Unbund DNA fragments are remved using 1x Wash Buffer Drying f VisinArray Chip by centrifugatin Duratin: 2 min Detectin Marking f bitinylated sequences using the Detectin Slutin Visualizatin is perfrmed by applying the Blue Spt Slutin 50 Min. Duratin: 17 min Wash Remving f the Blue Spt Slutin by washing with 1x Wash Buffer Drying f VisinArray Chip by centrifugatin Duratin: 2 min Analysis 60 Min. Chips are scanned with VisinArray Scanner 8100 Autmated analysis is perfrmed by using the VisinArray Analyzer Sftware Duratin: 10 min Quick & easy 1 hur prtcl Autmated user-friendly evaluatin within a few minutes

HPV Tests in Cmparisn Assay Gentyping C-infectin N.

250 CLART HPV (3, 4) 35 10 HPV 16 + 18 HPV 16 + 18 14 150 cpies/ml digene HC2 HPV DNA Test 18 3100 INNO-LiPA HPV Gentyping

a Glance Simultaneus gentyping f 41 different HPV types all certified fr in vitr diagnstic use All capture sequences and

5 HPV Tests in Cmparisn Assay Gentyping C-infectin N. f CE-IVD HPV Types (cpies/reactin) Anyplex TM II HPV28 Detectin Cervista HPV HR CLART HPV (3, 4) HPV HPV cpies/ml digene HC2 HPV DNA Test INNO-LiPA HPV Gentyping Extra II cbas HPV Test VisinArray * Sensitivity data are accrding t the respective instructin fr use VisinArray At a Glance Simultaneus gentyping f 41 different HPV types all certified fr in vitr diagnstic use All capture sequences and psitive cntrls are set up n the VisinArray Chip as duplicates High sensitivity and specificity Quick & easy 1 hur prtcl Autmated evaluatin using the VisinArray Analyzer Sftware simple visualizatin & quick analysis in just a few minutes Sensitivity*

VisinArray Arrays fr DNA analysis VisinArray HPV Chips Prd. N.

10 pieces VA-0001-50 VisinArray HPV Chip 1.0 50 Incl.

N. Prduct Tests VI-0001-50 VisinArray FFPE DNA Extractin Kit

Paraffin Disslver; Tissue Lysis Buffer; Decrsslink Buffer;

Buffer; Clumns; Cllectin Tubes VI-0002-50 VisinArray Cytlgy

Mix; dntp/dutp Slutin VE-0001-100 VisinArray PreCise Taq DNA

VisinArray PreCise Taq DNA Plymerase, Reactin Buffer, 10x;

10x ES-0007-50 VisinArray HPV PreCise Master Mix 50 Prd. N.

Slutin, 1 ml; Detectin Slutin, 5 ml; Blue Spt Slutin, 5 ml;

Scanner 8100; Slide Hlder; Hand Scanner; PC with preinstalled

Cmputer Muse nly available in certain cuntries.

6 VisinArray Arrays fr DNA analysis VisinArray HPV Chips Prd. N. Prduct Tests VA VisinArray HPV Chip Incl. 10 pieces VA VisinArray HPV Chip Incl. 5x 10 pieces VA VisinArray HPV High Risk Chip Incl. 10 pieces VA VisinArray HPV High Risk Chip Incl. 5x 10 pieces VisinArray DNA Extractin, PCR, and Detectin Prd. N. Prduct Tests VI VisinArray FFPE DNA Extractin Kit 50 Incl. Paraffin Disslver; Tissue Lysis Buffer; Decrsslink Buffer; DNA Wash Buffer; Prteinase K; Prteinase K Buffer; Elutin Buffer; Clumns; Cllectin Tubes VI VisinArray Cytlgy DNA Extractin Kit 50 Incl. Pre-Lysis Buffer; Cell Lysis Buffer; DNA Wash Buffer; Prteinase K; Prteinase K Buffer; Elutin Buffer; Clumns; Cllectin Tubes Prd. N. Prduct Tests VP VisinArray HPV Primer Kit 50 Incl. HPV Primer Mix; dntp/dutp Slutin VE VisinArray PreCise Taq DNA Plymerase 100 Incl. VisinArray PreCise Taq DNA Plymerase, Reactin Buffer, 10x; PreCise MgCl 2, 25 mm VE VisinArray Uracil-DNA Glycsylase 100 Incl. VisinArray Uracil- DNA Glycsylase (UDG), UDG Reactin Buffer, 10x ES VisinArray HPV PreCise Master Mix 50 Prd. N. Prduct Tests VK VisinArray Detectin Kit 50 Incl. Hybridizatin Slutin, 1 ml; Detectin Slutin, 5 ml; Blue Spt Slutin, 5 ml; 100x Wash Buffer, 250 ml E Mini Slide Centrifuge VisinArray Analysis Package Prd. N. E Prduct VisinArray Analysis Package Incl. Scanner 8100; Slide Hlder; Hand Scanner; PC with preinstalled VisinArray Analyzer Sftware; USB-Hub; External Hard Drive; Cmputer Muse nly available in certain cuntries. All ther cuntries research use nly! Please cntact yur lcal dealer fr mre infrmatin. M Manufactured by: 42 life sciences GmbH & C. KG Fischkai Bremerhaven Germany Distributed by: ZytVisin GmbH Fischkai Bremerhaven Germany inf@zytvisin.cm VisinArray is a trademark f 42 life sciences GmbH & C. KG

HIV Diagnostic Tests. HIV Testing Algorithm at SydPath (National Reference Laboratory)

HIV Diagnostic Tests. HIV Testing Algorithm at SydPath (National Reference Laboratory) HIV Diagnstic Tests HIV Testing Algrithm at SydPath (Natinal Reference Labratry) HIV1/2 Ab/Ag Cmbi is Architect HIV- 1 Ab/Ag EIA is Genscreen Sandwich EIA 4 th Generatin HIV Ab/Ag Chemiluminescene Micr