2019年北京市成人散发性腹泻病毒病原的基因特征研究

Study on the genetic characteristics of enteric viral pathogens of sporadic adult diarrhea in Chaoyang district,Beijing in 2019

目的分析2019年北京市成人散发性腹泻病毒病原的检出情况及其基因特征。方法收集2019年5—12月北京市朝阳区4家肠道门诊送检的177例成人散发性腹泻粪便样本177份,应用实时荧光定量PCR技术对样本中的诺如病毒、札如病毒、轮状病毒、肠道腺病毒和星状病毒进行核酸检测,对阳性样本利用普通PCR技术进行病毒核酸扩增并测序,采用Mega 6.0软件的Neighbor-Joining法进行系统进化分析。结果177份成人散发性腹泻样本中检测出60份病毒病原阳性(33.90%),其中47份检出单一病原,分别为诺如病毒29例、札如病毒9例、星状病毒8例和肠道腺病毒1例;13例患者样本检出两种病原;未检出轮状病毒。测序结果显示,诺如病毒GⅠ型测序成功16例,含7种基因亚型,主要为GⅠ.3[P13]亚型;诺如病毒GⅡ型测序成功10例,含5种基因亚型,以GⅡ.6[P7]亚型为主;札如病毒测序成功12例,含4种基因亚型,以GⅠ.2亚型为主;星状病毒测序成功7例,含2种基因亚型,以Ast-1亚型为主。结论2019年北京市成人散发性腹泻病毒病原以诺如病毒、星状病毒和札如病毒为主,不同病原基因亚型呈现出多样性特征。

Objective To analyze the distribution and genetic characteristics of sporadic adult diarrhea virus in Chaoyang District,Beijing.Methods Fecal samples from 177 adult patients with sporadic diarrhea were collected from 4 enteric outpatient clinics in Chaoyang District,Beijing from May to December 2019.Nucleic acid detection of Norovirus,Sappovirus,Rotavirus,Enteric Adenovirus and Astrovirus in the samples was performed by real-time quantitative PCR.The positive samples were amplified by RT-PCR/PCR and sequenced.The phylogenetic analysis was performed by neighbor-Joining(NJ)methods of Mega 6.0 software.Results There were 60 of 177(33.90%)adult sporadic diarrhea samples positive for enteric viral pathogens.Among them,47 cases were infected with single virus,including 29 cases of Norovirus,9 cases of Sappovirus,8 cases of Astrovirus and 1 case of Enteric Adenovirus,in addition with 13 cases of multiple infections.None of rotavirus was detected.Partial sequences were successfully obtained for analysis,including 16 cases of GI Norovirus(7 subtypes and GI.3[P13]predominant),10 cases of GII Norovirus(5 subtypes and GII.6[P7]predominant),12 cases of Sappovirus(4 subtypes and GI.2 predominant),and 7 cases of Astrovirus(2 subtypes and AST-1 predominant).Conclusion Norovirus,Astrovirus and Sappovirus are main pathogens among sporadic adult diarrhea in Beijing in 2019,and and different pathogenic gene subtypes show diverse characteristics.