Detection of bovine viral diarrhea virus, using degenerate oligonucleotide primers and the polymerase chain reaction.
A technique for detection of bovine viral diarrhea virus (BVDV) from circulating blood leukocytes, using the polymerase chain reaction, is described. The published nucleotide sequences of 2 strains of BVDV and that of hog cholera virus were aligned and the information was used to design oligonucleotides coding for 2 regions of amino acid homology. The oligonucleotides were a mixed population including all possible codons for the conserved amino acids. These degenerate oligonucleotides were used in the polymerase chain reaction to detect viral RNA in cells infected in vitro, or in circulating blood leukocytes from infected animals. Virus was detected in over 60 samples from diverse isolates. The detection of BVDV by the polymerase chain reaction is a rapid, sensitive, and specific technique, which represents an improvement over existing technology.
Ward
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P
Department of Veterinary Microbiology and BIOdiagnosTEC, Western College of Veterinary Medicine, Saskatoon, Saskatchewan, Canada.
Misra
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V
D000034:Abortion, Veterinary / Q000175:diagnosis; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001912:Bovine Virus Diarrhea-Mucosal Disease / Q000175:diagnosis* / Q000382:microbiology; D002417:Cattle; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000032:analysis* / Q000737:chemistry; D001908:Diarrhea Viruses, Bovine Viral / Q000235:genetics / Q000302:isolation & purification*; D005260:Female; D007962:Leukocytes / Q000382:microbiology; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000737:chemistry; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D011247:Pregnancy; D012367:RNA, Viral / Q000032:analysis* / Q000737:chemistry; D015183:Restriction Mapping; D012984:Software
D004279:DNA, Viral; D009841:Oligonucleotides; D012367:RNA, Viral