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Effect of carryover and presampling procedures on the results of real-time PCR used for diagnosis of bovine intramammary infections with Streptococcus agalactiae at routine milk recordings
Authors:Yasser S Mahmmod  Marshal M Mweu  Søren S Nielsen  Jørgen Katholm  Ilka C Klaas
Institution:1. HERD: Centre for Herd-oriented Education, Research and Development, Department of Large Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Grønnegårdsvej 2, DK-1870 Frederiksberg C, Denmark;2. Infectious Diseases, Department of Animal Medicine, Faculty of Veterinary Medicine, Zagazig University, Zagazig city 44511, Sharkia Province, Egypt;3. Department of Large Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Grønnegårdsvej 8, DK-1870 Frederiksberg C, Denmark;4. Knowledge Centre for Agriculture, Cattle, Agro Food Park 15, Skejby, DK-8200 Aarhus N, Denmark
Abstract:The use of PCR tests as diagnostics for intramammary infections (IMI) based on composite milk samples collected in a non-sterile manner at milk recordings is increasing. Carryover of sample material between cows and non-aseptic PCR sampling may be incriminated for misclassification of IMI with Streptococcus agalactiae (S. agalactiae) in dairy herds with conventional milking parlours. Misclassification may result in unnecessary costs for treatment and culling. The objectives of this study were to (1) determine the effect of carryover on PCR-positivity for S. agalactiae at different PCR cycle threshold (Ct) cut-offs by estimating the between-cow correlation while accounting for the milking order, and (2) evaluate the effect of aseptic presampling procedures (PSP) on PCR-positivity at the different Ct-value cut-offs.
Keywords:Carryover  Milking order correlation  PathoProof&trade  PCR  Presampling preparations  Teat disinfection  Test misclassification
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