Reid S M, Hutchings G H, Ferris N P, De Clercq K
Institute for Animal Health, Pirbright, Woking, Surrey, UK.
J Virol Methods. 1999 Dec;83(1-2):113-23. doi: 10.1016/s0166-0934(99)00113-5.
Multiple primers designed from the 1D and 2AB regions of the foot-and-mouth disease (FMD) viral genome were evaluated extensively for the detection of all seven serotypes of the virus by reverse transcription polymerase chain reaction (RT-PCR) at the OIE/FAO World Reference Laboratory for FMD (WRL), Pirbright. The primers had been characterised previously elsewhere on a relatively small number of cell culture grown isolates and epithelial suspensions and had been shown to identify and differentiate all seven serotypes of FMD virus. The extended study evaluated several RT-PCR protocols on epithelial suspensions and supernatant fluids, resulting from their passage in cell culture, derived from clinical samples of diverse molecular characteristics. Each of the serotype-specific primers in selected RT-PCR protocols demonstrated suitable specificity and detected cell culture passaged isolates with some success but were not adequate for the serotyping of suspensions prepared from clinical samples of epithelium. The results showed that the primers can be used in RT-PCR procedures in conjunction with the routine detection methods of virus isolation and ELISA for the diagnosis and serotyping of FMD virus.
在位于皮尔布赖特的世界动物卫生组织/联合国粮食及农业组织口蹄疫世界参考实验室（WRL），对从口蹄疫（FMD）病毒基因组的1D和2AB区域设计的多种引物进行了广泛评估，以通过逆转录聚合酶链反应（RT-PCR）检测该病毒的所有七种血清型。这些引物先前已在其他地方对数量相对较少的细胞培养物中生长的分离株和上皮悬浮液进行了表征，并已显示能够识别和区分口蹄疫病毒的所有七种血清型。这项扩展研究评估了几种针对上皮悬浮液和细胞培养传代后的上清液的RT-PCR方案，这些样本来自具有不同分子特征的临床样本。所选RT-PCR方案中的每种血清型特异性引物都表现出适当的特异性，并且在检测细胞培养传代分离株方面取得了一定成功，但不足以对从上皮临床样本制备的悬浮液进行血清分型。结果表明，这些引物可用于RT-PCR程序，结合病毒分离和ELISA等常规检测方法，对口蹄疫病毒进行诊断和血清分型。
