基于聚合酶区和衣壳区测序的诺如病毒暴发疫情溯源  

Tracing Norovirus outbreak based on sequencing of regions coding for capsid proteins and RNA polymerase

作　　者：何娇明 俞慕华[1] 陈辉[1] 马艳萍 张海龙[2] 鞠长燕[1] HE Jiao-ming;YU Mu-hua;CHEN Hui;MA Yan-ping;ZHANG Hai-long;JU Chang-yan(Nanshan Center for Disease Control and Prevention of Shenzhen,Shenzhen,Guangdong 518054,China;Shenzhen Center for Disease Control and Prevention,Shenzhen,Guangdong 518055,China)

机构地区：[1]深圳市南山区疾病预防控制中心,广东深圳518054 [2]深圳市疾病预防控制中心,广东深圳518055

出　　处：《中国热带医学》2022年第1期63-67,共5页China Tropical Medicine

基　　金：深圳市“医疗卫生三名工程”引进高层次医学团队项目(No.SZSM201803081);深圳市南山区医学重点学科项目。

摘　　要：目的分析一起厨工相关诺如病毒暴发的病原学特征,为诺如病毒感染的溯源提供证据。方法利用Filmarray病原体检测系统初筛,根据感染性腹泻诊断标准WS 271-2007进行细菌分离,同时进行诺如病毒荧光PCR检测,对诺如病毒阳性核酸进行聚合酶区和衣壳区序列扩增,测序比对,利用Mega软件分析同源性。结果 49份患者标本中,诺如GⅡ型阳性率为48.98%(24/49),GⅠ型阳性率为2.04%(1/49),GⅠ、GⅡ混合阳性率为2.04%(1/49);115份厨工及餐饮工作人员标本中,GⅡ型阳性率为13.04%(15/115),GⅠ型阳性率为0.87%(1/115)。38份环境涂抹样中,GⅡ型阳性率为2.63%(1/38)。有17份标本诺如病毒聚合酶区和衣壳区测序成功,其中患者GⅡ型14份,GⅠ型1份,厨工GⅡ型2份。比对结果显示1份GⅠ亚型为GⅠ.P4-GⅠ.5,16份GⅡ亚型均为GⅡ.P7-GⅡ.6。两份厨工和14份患者标本的诺如病毒核酸序列同源性达99%。49份患者标本中,致泻性大肠埃希菌毒力基因阳性率为34.69%(17/49),患者标本中合并诺如病毒核酸阳性和致泻性大肠埃希菌毒力基因阳性的有5份,阳性率为10.20%(5/49)。16份诺如病毒核酸阳性的厨工标本中,均未检出致泻性大肠埃希菌毒力基因,其余样本未进行致泻性大肠埃希菌检测。结论该起疫情是由诺如病毒GⅡ.P7-GⅡ.6亚型引起,患者和厨工极有可能来自同一传染源,同时合并致泻性大肠埃希菌感染,应加强厨工及餐饮服务人员的体检和培训。Objective To analyze the etiological characteristics of an outbreak of Norovirus related to kitchen workers, in order to provide evidence for tracing norovirus infection.Methods The pathogen detection system of Filmarray was used for preliminary screening. Bacteria were isolated according to the diagnostic standard of infectious diarrhea WS 271-2007.Norovirus was also detected by fluorescent PCR. The regions coding for capsid proteins and RNA polymerase were amplified,sequenced and compared during Norovirus positive nucleic acids, and the homology was analyzed by Mega software.Results Among 49 patients, the positive rate of norovirus G Ⅱ was 48.98%(24/49), G Ⅰ was 2.04%(1/49), and the mixed positive rate of GⅠand GⅡ was 2.04%(1/49). Among 115 samples of kitchen workers and related staff, the positive rate of Norovirus GⅡ was 13.04%(15/115), GⅠ was 0.87%(1/115). In 38 environmental samples, the positive rate of Norovirus GⅡwas 2.63%(1/38). The regions of Norovirus coding for capsid proteins and RNA polymerase was sequenced successfully in 17 samples, including 14 patients with GⅡtype, 1 patient with GⅠ type and 2 kitchen workers with GⅡ type. GⅠ subtype was GⅠ.P4-GⅠ.5, GⅡ subtype was GⅡ.P7-GⅡ.6. The norovirus sequences of 2 Kitchen workers and 14 patients were 99%homologous. Among 49 patients, the positive rate of virulence gene of diarrheal Escherichia coli was 34.69%(17/49). There were 5 patients mix infected Norovirus and virulence gene of diarrheal Escherichia coli, the positive rate was 10.20%(5/49). In16 positive norovirus samples of kitchen workers, the virulence gene of diarrheal Escherichia coli was not detected. The other samples were not tested for diarrheal Escherichia coli.Conclusions The outbreak is caused by norovirus GⅡ.P7-GⅡ.6 subtype. The patients and kitchen workers are likely to come from the same source of infection and are mix infected with diarrheal Escherichia coli. We should strengthen the physical examination and training of kitchen workers and related

关 键 词：急性胃肠炎 诺如病毒 混合感染 基因型 

分 类 号：R512.5[医药卫生—内科学]