Pattnaik B, Sanyal A, George M, Tosh C, Hemadri D, Venkataramanan R
Central Laboratory, Indian Veterinary Research Institute, Nainital, India.
Acta Virol. 1997 Dec;41(6):333-6.
Eight oligonucleotide primers in 7 different combinations were used to amplify 3D gene sequences of foot-and-mouth disease virus (FMDV) by reverse transcription-polymerase chain reaction (RT-PCR). Six of the primers were designed at this laboratory. All the primer combinations could specifically amplify 3D gene sequences of FMDV serotypes O, A, and C. The largest fragment amplified was of 1,393 bp and the smallest was of 208 bp in size. The 1,393 bp fragment included sequences from the preceeding P18 region of FMDV genome. The second largest fragment of 734 bp included sequences from the 3'-extracistronic region of viral genome. The remaining fragments were amplified either from the 3'- or 5'-half of the 3D gene. Specific amplification of the entire 3D gene in fragments of different size showed sequence conservation in the 3D genomic region of FMDV and usefulness of the primers reported in detecting inapparent or persistent FMDV infection in susceptible animals by RT-PCR.
使用7种不同组合的8条寡核苷酸引物，通过逆转录聚合酶链反应（RT-PCR）扩增口蹄疫病毒（FMDV）的3D基因序列。其中6条引物为本实验室设计。所有引物组合均可特异性扩增FMDV O、A和C型的3D基因序列。扩增出的片段中，最大的为1393 bp，最小的为208 bp。1393 bp的片段包含FMDV基因组中P18区域之前的序列。第二大的734 bp片段包含病毒基因组3'-非编码区的序列。其余片段则从3D基因的3'-或5'-半区扩增得到。不同大小片段中整个3D基因的特异性扩增表明，FMDV 3D基因组区域存在序列保守性，且所报道的引物可用于通过RT-PCR检测易感动物中不显性或持续性FMDV感染。
