The purpose of this study was to determine whether manually plucked hairs might serve as an alternative sample for a quantitative real time polymerase chain reaction (qRT-PCR) testing. Twenty three, 1~3 week old, non-bovine viral diarrhea virus (BVDV) vaccinated calves, found to be positive for BVDV by immunohistochemical staining, were selected and hairs were manually plucked from the ear. qRT-PCR was performed on samples consisting of more than 30 hairs (30~100) and whole blood. All 23 animals were positive for the virus by qRT-PCR performed on the whole blood and when samples of more than 30 hairs were assayed. Additionally, qRT-PCR was performed on groups of 10 and 20 hairs harvested from 7 out of 23 immunohistochemical staining-positive calves. When groups of 20 and 10 hairs were tested, 6 and 4 animals, respectively, were positive for the virus.
Animals ; Antibodies, Viral/analysis/diagnostic use ; Bovine Virus Diarrhea-Mucosal Disease/blood/*diagnosis/virology ; Cattle ; Diarrhea Virus 1, Bovine Viral/genetics/*isolation & purification ; Diarrhea Virus 2, Bovine Viral/genetics/*isolation & purification ; Hair/virology ; RNA, Viral/chemistry/genetics ; Real-Time Polymerase Chain Reaction/methods/*veterinary

Bovine viral diarrhea virus (BVDV) is a member of the genus Pestivirus, which is a widespread problem for beef and dairy herds, and has given rise to a significant loss in the livestock industry all over the world. The BVDV strain JZ05-1 isolated from cattle in Jilin, China generated cytopathic effect (CPE) in MDBK cells. Eight overlapped gene fragments were amplified by RT-PCR and sequenced, the complete genom sequence of BVDV strain JZ05-1 was assembled. According to the results, the JZ05-1 genome was composed of 12285 nucleotides in length (GenBank accession No. GQ888686), which could be divided into three regions: a 387 nt 5'-untranslated region (UTR), a 11694 nt single large open reading frame encoding a polyprotein, and a 204 nt 3'-UTR. The 5'-UTR and genome sequences were analyzed by sequence alignment and construction of phylogenetic trees. The strain JZ05-1 was classified as BVDV type 2a. The BVDV-2 strain JZ05-1 genome showed high similarity to the p11Q isolated in Canada and the XJ-04 isolated in China, with 90% and 91% identity in nucleotide sequence, respectively. Compared with the similarity within the BVDV-2 genotype (96%), the JZ05-1 had low sequence similarity to other BVDV-2 strains.
5' Untranslated Regions ; genetics ; Animals ; Cattle ; China ; Diarrhea Virus 2, Bovine Viral ; classification ; genetics ; isolation & purification ; Genome, Viral ; genetics ; Genotype ; Molecular Sequence Data ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA