Presentation is loading. Please wait.

Presentation is loading. Please wait.

Qualification Panels for West Nile Virus RNA Tests Mark Manak, Ph.D. Boston Biomedica, Inc. West Bridgewater, MA SoGAT Langen, Germany July 3, 2003.

Published byDennis Franklin Modified over 8 years ago

Similar presentations

Presentation on theme: "Qualification Panels for West Nile Virus RNA Tests Mark Manak, Ph.D. Boston Biomedica, Inc. West Bridgewater, MA SoGAT Langen, Germany July 3, 2003."— Presentation transcript:

1 Qualification Panels for West Nile Virus RNA Tests Mark Manak, Ph.D. Boston Biomedica, Inc. West Bridgewater, MA SoGAT Langen, Germany July 3, 2003

2 WNV Controls/Panels Panel/Controls Design Designed to closely mimic a true serum sample Cultured Virus from an infected individual Heat Treated Diluted in Defibrinated Plasma (Basematrix ) Panel/Controls Uses Method optimization and comparison Standardization of WNV RNA concentration Useful for quantitation Monitoring assay performance

3 West Nile Virus Qualification Panels QWN702 - WNV Lineage 1, NY 1999 (flamingo CDC) QWN701 - WNV Lineage 2, Uganda 1937 15 Member Panel 2 10,000 copies/ml 3 100 copies/ml 2 1,000 copies/ml 3 30 copies/ml 2 300 copies/ml 3 Negative Randomly Assorted Semi blinded

4 WNV Run Control: Accurun 365 Accurun 365 WNV Positive Run Control Manufactured from cultured West Nile Virus* Target level of 300 copies/ml Ideally suited to monitor assay precision Validation of analytical runs Run Control for Blood Screening Assays Accurun 865 WNV Negative Run Control * NY99 isolate, Lineage 2

5 RT-PCR Quantitative WNV Assay Specific Primers for WNV 5’UTR: Detects all known isolates of WNV Env: Lineage 1 only, more sensitive Closed-tube, Real-time fluorescent assay Quantified with TaqMan probe Broad linear dynamic range Between 300 – 10,000,000 copies/ml Ultrasensitive assay Sensitivity to 30 copies/ml

6 WNV RNA Standards-RNA Transcripts

7 WNV RNA Quantitation Standard Ultra 1,000 300 10,000 1,000 30 300 100 Copies/ml 30 Neg.

8 Reproducibility of Ultrasensitive Assay 100 30 Ave. Ct 32.2 35.0 STD 0.43 0.8 %CV 1.35 2.3 (n = 12)

9 QWN702 Panel Quantitation Panel MemberTarget 5’-LTR 140uL Env Primer 140uL Env CDC BBI Ultra Assay Ct QWN 701. 0310,00033.3629.2429.2526.32 QWN 701. 1210,00033.4329.1029.0526.00 QWN 701. 051,00036.6632.9632.0030.09 QWN 701. 101,00037.3532.7631.5529.91 QWN 701. 0630038.0432.9233.8030.36 QWN 701. 1530037.9434.1233.1030.50 QWN 701. 0110039.4933.2136.0532.69 QWN 701. 0710040.6035.9736.3032.21 QWN 701. 1110040.5035.4336.4032.88 QWN 701. 04305036.1439.1535.47 QWN 701. 093050 45.9034.61 QWN 701. 143050 44.6535.33 QWN 701. 02050 QWN 701. 08050 QWN 701. 13050

10 Comparison of Assays

11 Negative Specimens Material Source of materials No. of samples CtResults Serum American Red Cross 12850Negative Plasma BBI Diagnostics 10050Negative Basematrix BBI Diagnostics 9250Negative

12 WNV Positive Samples Target 10 8 10 7 10 6 10 5 10 4 10 3 Average (Copy/mL) 1.1x10 8 2.0x 10 7 2.1x 10 6 2.1x 10 5 2.9x 10 4 3.4x 10 3 STDEV 2.1x 10 7 5.7x 10 6 6.5x 10 5 3.9x 10 4 2.2x 10 4 2.6x 10 3 CV 19.0%28.5%31.3%18.6%78%76.2% #Samples 81312181410

13 WNV Titers in Avian and Mosquito Tissues Avian Brains Mosquito Pools ID copies/g ID copies/ml ZL02-249 0 SU02-01735 0 ZL02-251 0 SL02-01742 0 ZL02-255 0 SL02-01782 1,360 ZL02-257 0 EM02-04183 0 ZL02-265 0 NF02-07432 155,000 ZL02-754 136,000,000 NF02-07465 559,000 ZL02-755 37,800,000 NF02-07483 4,200,000 ZL02-756 52,200,000 SU02-10395 0 ZL02-760 10,400,000 SU02-10402 0 ZL02-761 190,000,000 SU02-10466 465,000

14 BBI WNV Panel and Controls BBI Panels and Controls –ACCURUN WNV RNA Positive Control A365 (NY99 Strain) –ACCURUN WNV RNA Negative Control A865 –QWN702 WNV Qualification Panel (Lineage 1) (NY99 Strain) –QWN701 WNV Qualification Panel (Lineage 2) (Uganda Strain) –WNV Characterized Samples (US Source ) Panels and controls Uses –Assay Development –Comparison/standardization of Assays –Training of Laboratory Personnel –Assay Validation –Monitoring Assay Performance

15 Acknowledgements Boston Biomedica Inc. Alan Doty Barbara Weiblen Harmesh Sharma American Red Cross Chyang Fang, Ph.D. MA State Health Depart. Barbara Werner, Ph.D. FDA Maria Rios, Ph.D. CDC Robert Lanciotti, Ph.D. BBI Biotech Xiuli Chen, Ph.D. Jienli Wu Ron Meixell

Download ppt "Qualification Panels for West Nile Virus RNA Tests Mark Manak, Ph.D. Boston Biomedica, Inc. West Bridgewater, MA SoGAT Langen, Germany July 3, 2003."

Similar presentations

Development of a Panel for Dengue Virus Maria Rios, PhD CBER/FDA Blood Products Advisory Committee Meeting December 14, 2010.

Development of a Panel for Dengue Virus Maria Rios, PhD CBER/FDA Blood Products Advisory Committee Meeting December 14, 2010.
Measuring the latent HIV Reservoir

Measuring the latent HIV Reservoir
Utilizing a Non-Commercial Real- Time PCR to Detect HIV-1 RNA in HIV Antibody Negative Diagnostic Sera Submitted to The Maryland Public Health Laboratory.

Utilizing a Non-Commercial Real- Time PCR to Detect HIV-1 RNA in HIV Antibody Negative Diagnostic Sera Submitted to The Maryland Public Health Laboratory.
Pediatric Diagnosis of HIV-1 Infection Using Dried Blood Spots Chin-Yih Ou, PhD NCHSTP/DHAP Centers for Disease Control and Prevention.

Pediatric Diagnosis of HIV-1 Infection Using Dried Blood Spots Chin-Yih Ou, PhD NCHSTP/DHAP Centers for Disease Control and Prevention.
Michael Turell, Kirti Dave, Maria Mayda, Zahra Parker, Russell Coleman, and Daniel Strickman US Army Medical Research Institute of Infectious Diseases,

Michael Turell, Kirti Dave, Maria Mayda, Zahra Parker, Russell Coleman, and Daniel Strickman US Army Medical Research Institute of Infectious Diseases,
Validating 16 Member Pooled APTIMA® HIV-1 RNA testing Ethridge Steven F, Sullivan T, Bennett B, Parker M, Hanson D, Hilliard J, Hart C, Patel P Diagnostic.

Validating 16 Member Pooled APTIMA® HIV-1 RNA testing Ethridge Steven F, Sullivan T, Bennett B, Parker M, Hanson D, Hilliard J, Hart C, Patel P Diagnostic.
Comparing the New EIAs with Old Standbys: Florida Bureau of Laboratories Verification Data HIV Diagnostics: New Developments and Challenges Feb. 28, 2005.

Comparing the New EIAs with Old Standbys: Florida Bureau of Laboratories Verification Data HIV Diagnostics: New Developments and Challenges Feb. 28, 2005.
Use of avidity reagent. Panbio Buffered Avidity Reagent Mild protein-denaturing solution that may be useful in differentiating recent infections from.

Use of avidity reagent. Panbio Buffered Avidity Reagent Mild protein-denaturing solution that may be useful in differentiating recent infections from.
Linearity Panels HIV RNA, HCV RNA, HBV DNA, and CMV DNA

Linearity Panels HIV RNA, HCV RNA, HBV DNA, and CMV DNA
BBI Biotech Research Laboratories Core B Janet L Lathey, Ph.D. Director Virology/Immunology.

BBI Biotech Research Laboratories Core B Janet L Lathey, Ph.D. Director Virology/Immunology.
Full characterization of HAV RNA window period positive blood donations in Germany SoGAT XVIII Bethesda.

Full characterization of HAV RNA window period positive blood donations in Germany SoGAT XVIII Bethesda.
R Diagnostics June 19, 2003 WNV FDA BPAC MeetingPCR NAT Blood Screening TaqScreen West Nile Virus Program - Roche Update James L. Gallarda, Ph.D. Roche.

R Diagnostics June 19, 2003 WNV FDA BPAC MeetingPCR NAT Blood Screening TaqScreen West Nile Virus Program - Roche Update James L. Gallarda, Ph.D. Roche.
Laboratory Investigation

Laboratory Investigation
TMA Assay for Detection of West Nile Virus BPAC Meeting - March 13, 2003 Cristina Giachetti, Ph.D. J.Linnen; A.Broulik; W. Wu; J.Cline; M.Lewis; G.Dennis;

TMA Assay for Detection of West Nile Virus BPAC Meeting - March 13, 2003 Cristina Giachetti, Ph.D. J.Linnen; A.Broulik; W. Wu; J.Cline; M.Lewis; G.Dennis;
HIV Testing Quality Assurance and Quality Control

HIV Testing Quality Assurance and Quality Control
PROFICIENCY TESTING OF IN-HOUSE NAT ASSAYS USED FOR BLOOD SCREENING XXI SoGAT International Working Group Meeting on the Standardization of NAT for the.

PROFICIENCY TESTING OF IN-HOUSE NAT ASSAYS USED FOR BLOOD SCREENING XXI SoGAT International Working Group Meeting on the Standardization of NAT for the.
E992750C 1 Replacement of HIV-1 p24 Antigen Screening with HIV-1 RNA Nucleic Acid Testing (NAT) for Whole Blood Donations S.L. Stramer, R.A. Porter, J.P.

E992750C 1 Replacement of HIV-1 p24 Antigen Screening with HIV-1 RNA Nucleic Acid Testing (NAT) for Whole Blood Donations S.L. Stramer, R.A. Porter, J.P.
Simoa Accelerator Laboratory

Simoa Accelerator Laboratory
HIV, HCV, and HBV NAT Controls Formulation, Stability and Performance Mark Manak BBI Diagnostics, Inc. A Division of SeraCare Life Sciences, Inc. SoGAT.

HIV, HCV, and HBV NAT Controls Formulation, Stability and Performance Mark Manak BBI Diagnostics, Inc. A Division of SeraCare Life Sciences, Inc. SoGAT.
Update on CBER HIV NAT panels and International panels Indira Hewlett, Ph.D Chief, Lab. of Molecular Virology DETTD/OBRR/FDA May 28, 2009 XXI SoGAT meeting.

Update on CBER HIV NAT panels and International panels Indira Hewlett, Ph.D Chief, Lab. of Molecular Virology DETTD/OBRR/FDA May 28, 2009 XXI SoGAT meeting.

Similar presentations

Ads by Google