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目的对2013年南京市疑似病毒性腹泻成人门诊样本进行星状病毒核酸检测、分子分型和进化分析,为星状病毒所致成人病毒性腹泻的分子流行病学提供资料。方法收集2013年疑似病毒性腹泻门诊成人病例的腹泻物375份,用星状病毒特异性引物和探针进行荧光定量RT-PCR检测,然后对荧光定量RT-PCR阳性的样本,用一步法RT-PCR再进行星状病毒特异片段的检测。对RT-PCR产物进行双向测序和进化树分析。结果荧光定量RT-PCR共检测到9例星状病毒阳性(2.4%),其中,6例(1.6%)扩增到单一条带,测序获得449 bp的星状病毒壳蛋白基因片段序列。经比对分析,6例星状病毒中,4例为5型星状病毒,1例为1型星状病毒,1例为8型星状病毒。结论南京市星状病毒所致成人病毒性腹泻中有多种型别星状病毒,5型为优势型。
OBJECTIVE To detect astrovirus nucleic acid, molecular typing and evolutionary analysis of adult outpatients suspected of viral diarrhea in Nanjing in 2013 to provide information on the molecular epidemiology of adult viral diarrhea caused by astrovirus. Methods A total of 375 diarrhea cases were collected from adult patients with suspected viral diarrhea in 2013. Real-time quantitative RT-PCR was performed using astrovirus-specific primers and probes. Then, one-step RT -PCR detection of astrovirus-specific fragments. RT-PCR products were subjected to bidirectional sequencing and phylogenetic analysis. Results A total of 9 astrocytomas (2.4%) were detected by RT-PCR, of which 6 (1.6%) were amplified into a single band and sequenced to obtain a 449 bp fragment of astrovirus capsid protein. Of the 6 astroviruses, 4 were type 5 astroviruses, 1 was type 1 astrovirus, and 1 was type 8 astrovirus. Conclusions There are many types of astroviruses in adult viral diarrhea caused by astrovirus in Nanjing, and type 5 is predominant.