RT-PCR and its detection limit for cell culture grown bluetongue virus 1 using NS1 gene group specific primers.
RT-PCR was standardised for the detection of bluetongue viral RNA using highly expressed non structural protein 1 gene as the target gene with specific primers targeted to 274 bp of 5' end of NS1 gene. PCR product was consistently obtained in 30 PCR cycles. Further, detection limit of RT-PCR was estimated using serial 10 fold dilutions of BHK 21 cells grown BTV 1. The study suggested that RT-PCR can be used for detection of BTV in Indian conditions with the sensitivity limit of 10 infectious particles of the virus. The study suggested that this technique may be used as a tool for sensitive detection of BTV in carrier/reservoir animals, insect vectors and certification of animals and their germ- plasm for export and import purposes.
Prasad
G
G
Department of Veterinary Microbiology, CCS Haryana Agricultural University, Hisar, India.
Minakshi
Malik
Y
Y
Maan
S
S
D000818:Animals; D001483:Base Sequence; D001820:Bluetongue virus / Q000235:genetics* / Q000302:isolation & purification; D002460:Cell Line; D006224:Cricetinae; D017931:DNA Primers / Q000235:genetics; D005814:Genes, Viral; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D017361:Viral Nonstructural Proteins / Q000235:genetics
D017931:DNA Primers; C078966:NS1 protein, bluetongue virus; D012367:RNA, Viral; D017361:Viral Nonstructural Proteins