一步RT-PCR方法在黑龙江省手足口病病原检测中的应用

Application of One Step RT-PCR Method for Pathogen Detection of Hand-foot-mouth Disease in Heilongjiang Province

目的了解核酸检测方法在黑龙江省手足口病病原谱调查中的应用价值,为疾病防控提供依据。方法在全省范围内收集2009年和2010年疑似手足口病病人疱疹液、咽拭子或肛拭子标本,使用人肠道病毒通用引物、肠道病毒71型(EV71)引物和萨奇病毒A组16型(CVA16)引物,采用一步RT-PCR方法进行病原核酸检测,进而分析病原构成,并对部分核酸检测阳性标本进行病毒分离。结果 2009年检测标本985份,其中EV71阳性226份、CVA16阳性191份、EV71和CVA16合并阳性3份,其他肠道病毒阳性100份。2010年检测标本1696份,其中EV71阳性354份、CVA16阳性356份、其他肠道病毒阳性188份。两年标本肠道病毒阳性率平均为52.87%,其中EV71和CVA16占阳性标本中病原总数的79.69%。选取353份核酸检测阳性标本进行病毒分离,共获得毒株265株。结论一步RT-PCR方法用于手足口病病原检测稳定、特异、快捷、易于分型,EV71和CVA16是黑龙江省近两年手足口病的主要病原体。

Objective To understand the application value of one step RT-PCR method for pathogen detection of Hand-foot-mouth disease（HFMD） in Heilongjiang Province,and to guide the disease prevention and control.Methods The herpes fluid,throat swab or rectal swab specimens of some of the suspected HFMD cases were collected and detected by using the one step RT-PCR method with enerovirus universal primers,enterovirus 71（EV71） primers and coxsackievirus A16（CVA16） primers in Heilongjiang province between 2009 and 2010.Then the pathogen characteristic was analyzed and the pathogen isolation was done for some of the cases.Results There were 985 samples were detected in 2009,and 226 samples of which were EV71,191 samples were CVA16,3 samples were co-infected with EV71 and CAV16,100 samples were other enteroviruses infection.In 2010,1696 samples were detected,and 354 samples of which were EV71,356 samples were CVA16,188 samples were other enteroviruses infection.The average positive rate of enterovirus infection was 52.87% every year,and about 79.69% of the total of positive samples were EV71 or CVA16 infection.Then we selected 353 samples for virus isolation,and obtained 265 isolates finally.Conclusion One step RT-PCR method was stable,specific,convenient for direct identification and typing of HEV.EV71 and CVA16 were the major pathogens of HFMD in recent years.