title: Comparison of virus isolation and reverse transcription polymerase chain reaction assay for detection of bovine viral diarrhea virus in bulk milk tank samples.
abstract: The use of a reverse transcriptase polymerase chain reaction (RT-PCR) assay to screen bulk milk tank samples for bovine viral diarrhea virus (BVDV) has proven to be a sensitive and economical means to evaluate the lactating animals in a herd. The assay is capable of detecting the presence of a single 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 RT-PCR assay in conjunction with a classical virus isolation (VI) procedure to measure the relative effectiveness of the techniques. Virus could be detected with both methods when the milk from a single persistently infected animal 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 infection, and the correlation between the 2 assays was 95.9%. In terms of sensitivity, specificity, and turnaround time, RT-PCR was superior to VI. However, 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 contain minor sequence variations in the primer regions. VI and RT-PCR are both suitable for detection of BVDV in bulk milk samples when used independently, but to increase the probability of successful detection and to provide cross-checks against assay contamination, it is desirable to utilize both methods in parallel.
authors.affiliation: Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14850, USA.
authors.forename: R W
authors.initials: RW
authors.lastname: Renshaw
authors.forename: R
authors.initials: R
authors.lastname: Ray
authors.forename: E J
authors.initials: EJ
authors.lastname: Dubovi
mesh_terms: D000818:Animals; D001483:Base Sequence; D002417:Cattle; D004279:DNA, Viral / Q000032:analysis; D001908:Diarrhea Viruses, Bovine Viral / Q000302:isolation & purification*; D005188:False Negative Reactions; D005506:Food Contamination; D008892:Milk / Q000821:virology*; D008969:Molecular Sequence Data; D015995:Prevalence; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary*; D012680:Sensitivity and Specificity
chemical_list: D004279:DNA, Viral