双重逆转录PCR检测脊髓灰质炎病毒和肠道病毒

Double reverse transcription PCR detect poliovirus and enterovirus

目的 构建双重逆转录PCR对急性弛缓性麻痹(AFP)病例中脊髓灰质炎病毒(PV)和肠道病毒(EV)进行检测和基因分型。方法 从2021年河南省AFP病例中细胞分离得到的36株EV和7株PV粪便标本,利用EV VP4区引物EVP4、OL68-1和PV VP1区引物UG1、UC11构建双重逆转录PCR进行检测,分析特异性和敏感性;以细胞分离培养阳性结果为参照,分析双重逆转录PCR检测的灵敏性。结果 18株EV检测为阳性,4株EV检测为弱阳性,阳性率为61.1%;5株PV检测为阳性,阳性率为71.4%。EV以柯萨奇B组病毒3型为主,PV均为疫苗类似株。结论 本次实验构建的双重逆转录PCR可以作为PV和EV的辅助诊断手段。

Objective To construct double reverse transcription PCR for detecting and genotyping poliovirus and enterovirus from acute flaccid paralysis(AFP) cases. Methods Fecal specimens of 36 strains of EV and 7 strains of PV were isolated from AFP cases in in Henan province in 2021.The primers EVP4/OL68-1 of enterovirus VP4 region and primers UG1/UC11 of poliovirus VP1 region were used to detect the specificity and sensitivity;taking the positive results of cell isolation and culture as the reference, the sensitivity of double reverse transcription PCR was analyzed. Results A total of 18 strains of enterovirus were positive and 4 strains of enterovirus were weakly positive, with a positive rate of 61.1%;Five strains of poliovirus were positive,and the positive rate was 71.4%. Coxsackievirus B3 was the main enterovirus and poliovirus was vaccine strain. Conclusions The double reverse transcription PCR constructed in this experiment can be used as an auxiliary diagnostic method for poliovirus and enterovirus.