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目的了解龙岩市大量判定为其他肠道病毒(EV)感染的手足口病(HFMD)样本的病原组成。方法对龙岩市2011年经Real-time RT-PCR检测判为未分型EV感染的104份HFMD咽拭子样本经RD细胞分离培养后以RT-PCR法检测EV、EV71、CoxA16,对仍不能分型的样本以187/222引物进行扩增,对扩增产物进行纯化回收、克隆、测序,并用Blast程序在线比对判定型别。同时对培养物进行Real-time RT-PCR检测EV、EV71、CoxA16。任一法检出培养物阳性即判为阳性。结果 104份样本中检出EV阳性88份(84.62%),包括29份EV71,4份EV71+CoxA16,3份CoxB5,2份CoxA10,50份仍未能分型。结论 Real-time RT-PCR检测未能分型的HFMD病例中,相当部分(37.18%)还是由EV71所致,临床不应该随意排除该类病例EV71感染的可能,同时还包含CoxB5和CoxA10病毒。
Objective To understand the pathogenic composition of a large number of hand, foot and mouth disease (HFMD) samples in Longyan city, which were judged as other enterovirus (EV) infections. Methods Totally 104 HFMD throat swabs, which were classified as non-typed EVs by Real-time RT-PCR in 2011, were isolated and cultured in RD cells. EV, EV71 and CoxA16 were detected by RT-PCR. The genotyping samples were amplified with 187/222 primers. The amplified products were purified, recovered, cloned and sequenced. The genotypes were determined by Blast program online. Cultures were also subjected to Real-time RT-PCR to detect EV, EV71, CoxA16. Any method detected culture positive is judged as positive. Results 88 positive samples (84.62%) were detected in 104 samples, including 29 EV71, 4 EV71 + CoxA16, 3 copies CoxB5 and 2 copies CoxA10. Conclusion A significant proportion (37.18%) of HFMD cases that failed to be typed by Real-time RT-PCR were still caused by EV71. The clinical trial should not exclude the possibility of EV71 infection in this type of cases, as well as CoxB5 and CoxA10 viruses.