Chezzi C
Department of Virology, University of the Witwatersrand, Johannesburg, South Africa.
J Clin Microbiol. 1996 Jul;34(7):1722-5. doi: 10.1128/JCM.34.7.1722-1725.1996.
A single-tube, single-primer-set reverse transcription-PCR assay was developed for the rapid detection of polioviruses in infected tissue culture fluids and clinical materials. The poliovirus-specific PCR primers are located in the VP1-2A region of the poliovirus genome. They generate a 290-bp product and can be used in duplex reactions with general enterovirus primers. The primers span the region used for genotype determination, so that genotype analysis of wild-type polioviruses can be performed by direct sequencing of the PCR products. Of 125 virus isolates typed as polioviruses by neutralization assays, 125 (100%) were also positive by PCR, and of 38 isolates typed as non-polio enteroviruses by conventional techniques, 38 (100%) were also negative by PCR. The assay described here is rapid, highly sensitive, and specific and has clinical applicability in the diagnosis of poliovirus infections.
开发了一种单管、单引物组逆转录聚合酶链反应(RT-PCR)检测方法,用于快速检测感染的组织培养液和临床材料中的脊髓灰质炎病毒。脊髓灰质炎病毒特异性PCR引物位于脊髓灰质炎病毒基因组的VP1-2A区域。它们产生一个290 bp的产物,可用于与一般肠道病毒引物的双重反应。这些引物跨越用于基因型测定的区域,因此野生型脊髓灰质炎病毒的基因型分析可通过对PCR产物进行直接测序来进行。通过中和试验鉴定为脊髓灰质炎病毒的125株病毒分离株,PCR检测也均为阳性(100%);通过传统技术鉴定为非脊髓灰质炎肠道病毒的38株分离株,PCR检测也均为阴性(100%)。本文所述检测方法快速、高度灵敏且特异,在脊髓灰质炎病毒感染的诊断中具有临床适用性。