Ring test evaluation of the detection of influenza A virus in swine oral fluids by real-time reverse-transcription polymerase chain reaction and virus isolation

Date
2016-01-01
Authors
Goodell, Christa
Zhang, Jianqiang
Strait, Erin
Wang, Chong
Harmon, Karen
Patnayak, Devi
Otterson, Tracy
Culhane, Marie
Christopher-Hennings, Jane
Clement, Travis
Leslie-Steen, Pamela
Hesse, Richard
Anderson, Joe
Skarbek, Kevin
Vincent, Amy
Kitikoon, Pravina
Swenson, Sabrina
Jenkins-Moore, Melinda
McGill, Jodi
Rauh, Rolf
Nelson, William
O'Connell, Catherine
Shah, Rohn
Wang, Chong
Main, Rodger
Zimmerman, Jeffery
Major Professor
Advisor
Committee Member
Journal Title
Journal ISSN
Volume Title
Publisher
Altmetrics
Authors
Wang, Chong
Person
Research Projects
Organizational Units
Journal Issue
Series
Department
StatisticsVeterinary Diagnostic and Production Animal Medicine
Abstract

The probability of detecting influenza A virus (IAV) in oral fluid (OF) specimens was calculated for each of 13 assays based on real-time reverse-transcription polymerase chain reaction (rRT-PCR) and 7 assays based on virus isolation (VI). The OF specimens were inoculated with H1N1 or H3N2 IAV and serially diluted 10-fold (10(-1) to 10(-8)). Eight participating laboratories received 180 randomized OF samples (10 replicates × 8 dilutions × 2 IAV subtypes plus 20 IAV-negative samples) and performed the rRT-PCR and VI procedure(s) of their choice. Analysis of the results with a mixed-effect logistic-regression model identified dilution and assay as variables significant (P < 0.0001) for IAV detection in OF by rRT-PCR or VI. Virus subtype was not significant for IAV detection by either rRT-PCR (P = 0.457) or VI (P = 0.101). For rRT-PCR the cycle threshold (Ct) values increased consistently with dilution but varied widely. Therefore, it was not possible to predict VI success on the basis of Ct values. The success of VI was inversely related to the dilution of the sample; the assay was generally unsuccessful at lower virus concentrations. Successful swine health monitoring and disease surveillance require assays with consistent performance, but significant differences in reproducibility were observed among the assays evaluated.

Comments

This article is from The Canadian Journal of Veterinary Research 80 (2016); 12

Description
Keywords
Citation
DOI
Source
Collections