聚合酶链反应用于脊髓灰质炎病毒的诊断和定型

IDENTIFICATION OF POLlOVlRUS TYPES USING POLYMERASE CHAIN REACTION

利用聚合酶链反应(Polymerase chian reaction,PCR)对脊髓灰质炎病毒Sabin相关株进行了诊断和定型。选用的引物Map7、Map14与肠道病毒具有同源性,经扩增后产生大小为115bp的核酸片段;引物Sabin Ⅰ-1、Ⅰ-2对Ⅰ型Sabin株特异,经扩增后产生大小为97bp的核酸片段;引物Sabin Ⅱ-1、Ⅱ-2对Sabin Ⅱ型株特异,扩增产物为71bp;引物Sa-bin Ⅲ-1、Ⅲ-2对Sabin Ⅲ型株特异,扩增产物为44bp.此方法敏感性高,可检出10^(1.5)TCID_(50)/ml感染量的病毒核酸。利用PCR方法对我国各地送检的几株脊髓灰质炎病毒的定型结果,与中和试验完全一致。

This paper reports the identification of three types of poliovirus and Sabin-related strain by polymerase chain reaction ( PCR ) . Four pairs of synthetic oligonucleotide primers are used. The pair primer map 7 and map 14 which is homologous to all enterovirus can produce 115bp nucleic fragment by PCR amplification. Other three pairs ( Sabin I-1 and Sabin I-2, Sabin II-1 and Sabin II-2, Sabin III-1 and Sabin III-2) which are specific to Sabin I, Sabin I, and SabinI can produce 97bp, 7lbp and 44bp nucleic fragment by PCR respectively. The method is sensitive and TCID 50/ml 101.5 titre of virus can be detected by PAGE after amplification. Six isolates were examined by PCR using three pairs of specific primers. Of four strain which were isolated from Liaoning and Zhejiang Province, three were positive to Sabin I and one was positive to Sabin I. The result is accordance with neutralization and IgM test. Another two strains isolated from the poliomyelitis epidemic in Shandong Province were negative when the Sabin primers were used.