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Title:   Author  item
                                            GOODELL, CHRISTA - Iowa State Universityitem
                                            ZHANG, JIANQIANG - Iowa State Universityitem
                                            STRAIT, ERIN - Iowa State Universityitem
                                            HARMON, KAREN - Iowa State Universityitem
                                            PATNAYAK, DEVI - University Of Minnesotaitem
                                            OTTERSON, TRACY - University Of Minnesotaitem
                                            GRAMER, MARIE - University Of Minnesotaitem
                                            CHRISTOPHER-HENNINGS, JANE - South Dakota State Universityitem
                                            Baker, Amyitem
                                            Kitikoon, Pravinaitem
                                            McGill, Jodi
  Submitted to: Canadian Journal of Veterinary Research
                                    Publication Type: Peer Reviewed Journal
                                    Publication Acceptance Date: 5/19/2015
                                    Publication  Date: 1/16/2016
                                    Citation: Goodell, C.K., Zhang, J., Strait, E., Harmon, K., Patnayak, D., Otterson, T., Culhane, M., Christopher-Hennings, J., Clement, T., Leslie-Steen, P., Hesse, R., Anderson, J., Skarbek, K., Vincent, A., Kitikoon, P., Swenson, S., Jenkins-Moore, M., McGill, J., Rauh, R., Nelson, W., O'Connell, C., Shah, R., Wang, C., Main, R., Zimmerman, J.J. 2016. 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. Canadian Journal of Veterinary Research. 80(1):12-20.Interpretive Summary: Influenza A virus (IAV) is an important respiratory pathogen in pigs with occasional spread of swine IAV to humans. Rapid and reliable detection of IAV infections in pigs is necessary to prevent spread of the disease. Diagnostic samples that can be collected on live animals without restraint are preferred in many situations, and oral fluids (OF) are such a sampling option. This study was conducted to evaluate the performance of routine diagnostic tests (RT-PCR and virus isolation) for IAV among 8 laboratories on OF samples from swine spiked with known amounts of IAV. Successful swine health monitoring and disease surveillance require assays with consistent performance, but significant differences in reproducibility were observed among the laboratories for detecting IAV in OF, particularly as concentration of virus decreased. Further efforts are required to improve accurate and consistent performance of OF as a diagnostic specimen for IAV among veterinary laboratories.
                                Technical Abstract: The probability of detecting influenza A virus (IAV) in oral fluid (OF) specimens was calculated for each of 13 real-time, reverse transcription polymerase chain reaction (rRT-PCR) and 7 virus isolation (VI) assays. To conduct the study, OF was inoculated with H1N1 or H3N2 IAV and serially 10-fold diluted (10-1 to 10-8). Eight participating laboratories received 180 randomized samples (10 replicates x 8 dilutions x 2 IAV subtypes plus 20 IAV-negative oral fluid samples) and performed the IAV rRT-PCR and VI procedure(s) of their choice. Analysis of the results using a mixed-effect logistic regression model identified dilution (p <0.0001) and assay (p <0.0001) as variables significant to IAV detection in OF by rRT-PCR or VI. Virus subtype was not significant to IAV detection by either rRT-PCR (p = 0.4566) or VI (p = 0.1006). For rRT-PCRs, cycle threshold (Ct) values increased consistently with dilution, but exhibited wide variation. Therefore, it was not possible to predict VI success based on Ct values. VI was inversely related to the dilution of the sample and 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.


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