Comparison of virus isolation and reverse transcription polymerase chain reaction assay for detection of bovine viral diarrhea virus in bulk milk tank samples

The use of a reverse transcriptase polymerase chain reaction (RT-PCR) assay to screen bulk milk tank samples for bovine viral diarrhea virus (BVDV) ha s proven to be a sensitive and economical means to evaluate the lactating a nimals in a herd. The assay is capable of detecting the presence of a singl e persistently infected animal within a group of several hundred cows. Over a 3-year period, 144 samples from 97 farms were tested for BVDV using an R T-PCR assay in conjunction with a classical virus isolation (VI) procedure to measure the relative effectiveness of the techniques. Virus could be det ected with both methods when the milk from a single persistently infected a nimal was diluted 1:600 with the milk from a herd of BVDV-negative animals. Based on individual farms, there was an overall prevalence of 12.4% BVDV i nfection, and the correlation between the 2 assays was 95.9%. In terms of s ensitivity, specificity, and turnaround time, RT-PCR was superior to VI. Ho wever, of the 17 samples that were VI positive, 4 were RT-PCR negative. RT- PCR may not detect all naturally occurring BVDV isolates because they may c ontain minor sequence variations in the primer regions. VI and RT-PCR are b oth suitable for detection of BVDV in bulk milk samples when used independe ntly, but to increase the probability of successful detection and to provid e cross-checks against assay contamination, it is desirable to utilize both methods in parallel.