Comparison of Five Diagnostic Methods for Detecting Bovine Viral Diarrhea Virus Infection in Calves

Abstract

Abstract. Five diagnostic techniques performed on skin biopsies (shoulder region) and/or serum were compared for detection of bovine viral diarrhea virus infection in 224 calves 0-3 months of age, 23 calves older than 3 months but younger than 7 months, and 11 cattle older than 7 months. The diagnostic methods used were immunohistochemistry (IHC), 2 commercial antigen ELISAs, 1 commercial antibody ELISA, and realtime RT-PCR. Results of 249 out of 258 skin and serum samples were identical and correlated within the 3 antigen detection methods and the real-time RT-PCR used. Twenty-six of these 249 samples were BVDVpositive with all antigen detection methods and the real-time RT-PCR. Nine out of 258 samples yielding discordant results were additionally examined by RT-PCR, RT-PCR Reamplification (ReA), and antigen ELISA I on serum and by immunohistochemistry on formalin fixed and paraffin-embedded skin biopsies. Virus isolation and genotyping was performed as well on these discordant samples. In 3 cases, transiently infected animals were identified. Two samples positive by real-time RT-PCR were interpreted as false positive and were ascribed to cross-contamination. The antigen ELISA II failed to detect 2 BVDV-positive calves due to the presence of maternal antibodies; the cause of 2 false-positive cases in this ELISA remained undetermined. Only persistently infected animals were identified in skin samples by IHC or antigen ELISA I. The 3 antigen detection methods and the real-time RT-PCR used in parallel had a high correlation rate (96.5%) and similar sensitivity and specificity values.

BVD eradication in Switzerland—A new approach

“…The sample is sent to one of the nine certified laboratories involved in the program the same day. In the laboratories respectively 97.1% and 100% based on literature (Wolf et al, 2007;Fux, 2007;Fux et al, 2008;Hilbe et al, 2007).…”

“…The improvement of diagnostic tests for the detection of PI-animals in recent years in terms of early detection of young infected animals and the possibility to use ear-notch samples made the Swiss approach feasible (Hilbe et al, 2007;Houe et al, 2006). In 2008, all cattle will be tested.…”

Monitoring for Bovine Viral Diarrhea Virus in Austrian Red Deer (Cervus elaphus elaphus) by using Ear-notch Samples

“…Since 2007, this method of BVDV control has been applied to all offspring calves in the federal Austrian state of Tyrol. The diagnostic advantage of using skin samples instead of serum or plasma is that there is less interference with maternal antibodies at a very early stage when detecting PI animals (Hilbe et al, 2007). Moreover samples can be stored for a long time without losing the potential for BVDV antigen detection.…”

Genetic typing of bovine viral diarrhoea virus in cattle on Irish farms

“…Furthermore, Cornish et al (2005) revealed that the antigen capture ELISA method identified 100% of PI animals in the sample population analysed when compared to a number of other diagnostic techniques. Hilbe et al (2007) reported varying antigen ELISA sensitivity and specificity values in a diagnostic method comparison. The initial ELISA achieved 100% sensitivity and specificity values while analysis of the second assay's performance revealed 93.5% and 99% respectively as this method failed to detect two infected animals (Hilbe et al 2007).…”

“…Hilbe et al (2007) reported varying antigen ELISA sensitivity and specificity values in a diagnostic method comparison. The initial ELISA achieved 100% sensitivity and specificity values while analysis of the second assay's performance revealed 93.5% and 99% respectively as this method failed to detect two infected animals (Hilbe et al 2007). ELISA's that detect antibodies against BVDV proteins, such as the p80 and E 0 proteins, can be a useful screening tool as the status of a herd can be economically established through testing a bulk milk sample (Sayers et al 2015b) or pooled serum samples (Cowley et al 2012) samples to reduce cost (Dubovi 2013).…”