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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 value of nucleic acid detection in the investigation of pathogen spectrum of HFMD in Heilongjiang Province and provide the basis for disease prevention and control. Methods Herpes fluid, throat swabs or anal swab samples from suspected HFMD patients in 2009 and 2010 were collected across the province. Human enterovirus common primers, enterovirus 71 (EV71) primers and SSRV A group of 16 type (CVA16) primers, using one-step RT-PCR method of pathogenic nucleic acid detection, and then analysis of pathogenic constitution, and some of the nucleic acid detection of positive specimens for virus isolation. Results In 2009, 985 samples were detected, including 226 EV71 positives, 191 positive CVA16s, 3 positive EV71s and CVA16s, and 100 positive other enteroviruses. In 2010, 1696 samples were tested, of which 354 were EV71 positive, 356 were CVA16 positive and 188 were other enterovirus positive. The positive rate of enterovirus in two years was 52.87% on average, of which, EV71 and CVA16 accounted for 79.69% of the total number of pathogens in positive specimens. A total of 265 strains were obtained from 353 positive samples tested for virus isolation. Conclusion One-step RT-PCR method is used to detect pathogen of HFMD, which is stable, specific, rapid and easy to type. EV71 and CVA16 are major pathogens of HFMD in Heilongjiang Province in recent two years.