Diseases of Aquatic Organisms
DAO is a hybrid research journal on all aspects of disease phenomena in aquatic organisms.
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DOI: https://doi.org/10.3354/dao03687
copiedPerformance characteristics of two real-time TaqMan polymerase chain reaction assays for the detection of WSSV in clinically diseased and apparently healthy prawns
- Nicholas J. G. Moody
- Peter G. Mohr
- Lynette M. Williams
- David M. Cummins
- John Hoad
- J. Slater
- Stacey T. Valdeter
- Axel Colling
- Nagendrakumar B. Singanallur
- Ian A. Gardner
- Nicholas Gudkovs
- Mark St. J. Crane
ABSTRACT: This study aimed to generate data on performance characteristics for 2 real-time TaqMan PCR assays (CSIRO and WOAH WSSV qPCRs) for the purposes of (1) detection of white spot syndrome virus (WSSV) in clinically diseased prawns and (2) detection of WSSV in apparently healthy prawns. Analytical sensitivity of both assays was 2 to 20 genome copies per reaction, and analytical specificity was 100% after testing nucleic acid from 9 heterologous prawn pathogens and 4 prawn species. Results obtained after testing more than 20 000 samples in up to 559 runs with the CSIRO WSSV qPCR and up to 293 runs with the WOAH WSSV qPCR demonstrated satisfactory repeatability for both assays. Both assays demonstrated median diagnostic sensitivity (DSe) 100% (95% CI: 94.9-100%) when testing clinically diseased prawns. When 1591 test results from apparently healthy prawns were analysed by Bayesian latent class analysis, median DSe and diagnostic specificity (DSp) were 82.9% (95% probability interval [PI]: 75.0-90.2%) and 99.7% (95% PI: 98.6-99.99%) for the CSIRO WSSV qPCR and 76.8% (95% PI: 68.9-84.9%) and 99.7% (95% PI: 98.7-99.99%) for the WOAH WSSV qPCR. When both assays were interpreted in parallel, median DSe increased to 98.3 (95% PI: 91.6-99.99%), and median DSp decreased slightly to 99.4% (95% PI: 97.9-99.99%). Routine testing of quantified positive controls by laboratories in the Australian laboratory network demonstrated satisfactory reproducibility of the CSIRO WSSV qPCR assay. Both assays demonstrated comparable performance characteristics, and the results contribute to the validation data required in the WOAH validation pathway for the purposes of detection of WSSV in clinically diseased and apparently healthy prawns.
KEYWORDS
Nicholas J. G. Moody (Corresponding Author)
- CSIRO Australian Centre for Disease Preparedness, 5 Portarlington Rd, East Geelong, Victoria 3220, Australia
- WOAH Collaborating Centre for Diagnostic Test Validation Science in the Asia Pacific Region, 5 Portarlington Rd, East Geelong, Victoria 3220, Australia
Peter G. Mohr (Co-author)
- CSIRO Australian Centre for Disease Preparedness, 5 Portarlington Rd, East Geelong, Victoria 3220, Australia
- WOAH Collaborating Centre for Diagnostic Test Validation Science in the Asia Pacific Region, 5 Portarlington Rd, East Geelong, Victoria 3220, Australia
Lynette M. Williams (Co-author)
- CSIRO Australian Centre for Disease Preparedness, 5 Portarlington Rd, East Geelong, Victoria 3220, Australia
- WOAH Collaborating Centre for Diagnostic Test Validation Science in the Asia Pacific Region, 5 Portarlington Rd, East Geelong, Victoria 3220, Australia
David M. Cummins (Co-author)
- CSIRO Australian Centre for Disease Preparedness, 5 Portarlington Rd, East Geelong, Victoria 3220, Australia
- WOAH Collaborating Centre for Diagnostic Test Validation Science in the Asia Pacific Region, 5 Portarlington Rd, East Geelong, Victoria 3220, Australia
John Hoad (Co-author)
- CSIRO Australian Centre for Disease Preparedness, 5 Portarlington Rd, East Geelong, Victoria 3220, Australia
- WOAH Collaborating Centre for Diagnostic Test Validation Science in the Asia Pacific Region, 5 Portarlington Rd, East Geelong, Victoria 3220, Australia
J. Slater (Co-author)
- CSIRO Australian Centre for Disease Preparedness, 5 Portarlington Rd, East Geelong, Victoria 3220, Australia
- WOAH Collaborating Centre for Diagnostic Test Validation Science in the Asia Pacific Region, 5 Portarlington Rd, East Geelong, Victoria 3220, Australia
Stacey T. Valdeter (Co-author)
- CSIRO Australian Centre for Disease Preparedness, 5 Portarlington Rd, East Geelong, Victoria 3220, Australia
- WOAH Collaborating Centre for Diagnostic Test Validation Science in the Asia Pacific Region, 5 Portarlington Rd, East Geelong, Victoria 3220, Australia
Axel Colling (Co-author)
- CSIRO Australian Centre for Disease Preparedness, 5 Portarlington Rd, East Geelong, Victoria 3220, Australia
- WOAH Collaborating Centre for Diagnostic Test Validation Science in the Asia Pacific Region, 5 Portarlington Rd, East Geelong, Victoria 3220, Australia
Nagendrakumar B. Singanallur (Co-author)
- CSIRO Australian Centre for Disease Preparedness, 5 Portarlington Rd, East Geelong, Victoria 3220, Australia
- WOAH Collaborating Centre for Diagnostic Test Validation Science in the Asia Pacific Region, 5 Portarlington Rd, East Geelong, Victoria 3220, Australia
Ian A. Gardner (Co-author)
- WOAH Collaborating Centre for Diagnostic Test Validation Science in the Asia Pacific Region, 5 Portarlington Rd, East Geelong, Victoria 3220, Australia
- Department of Health Management, Atlantic Veterinary College, University of Prince Edward Island, 550 University Avenue, Charlottetown, PEI C1A 4P3, Canada
Nicholas Gudkovs (Co-author)
- CSIRO Australian Centre for Disease Preparedness, 5 Portarlington Rd, East Geelong, Victoria 3220, Australia
- WOAH Collaborating Centre for Diagnostic Test Validation Science in the Asia Pacific Region, 5 Portarlington Rd, East Geelong, Victoria 3220, Australia
Mark St. J. Crane (Co-author)
- CSIRO Australian Centre for Disease Preparedness, 5 Portarlington Rd, East Geelong, Victoria 3220, Australia
- WOAH Collaborating Centre for Diagnostic Test Validation Science in the Asia Pacific Region, 5 Portarlington Rd, East Geelong, Victoria 3220, Australia