上海单中心急性胃肠炎患儿感染诺如病毒的基因特征分析  

Genetic characteristics of norovirus in children with acute gastroenteritis in a single center in Shanghai

作　　者：于方圆 孟磊俊 潘芬[1] 马展[1] 王春[1] 孙燕[1] 周颖 张泓[1] Yu Fangyuan;Meng Leijun;Pan Fen;Ma Zhan;Wang Chun;Sun Yan;Zhou Ying;Zhang Hong(Department of Clinical Laboratory,Children′s Hospital of Shanghai,Children′s Hospital of Shanghai Jiaotong University,Shanghai 200040,China)

机构地区：[1]上海市儿童医院上海交通大学附属儿童医院检验科,200040

出　　处：《中华微生物学和免疫学杂志》2020年第10期792-799,共8页Chinese Journal of Microbiology and Immunology

基　　金：上海市重点临床专科建设项目(shslczdzk06902)。

摘　　要：目的了解上海地区急性胃肠炎患儿感染诺如病毒(norovirus,NoV)的基因特征情况。方法收集2018年10月至2019年9月上海市儿童医院门急诊急性胃肠炎患儿的粪便标本709份,采用实时荧光RT-PCR方法对NoV进行初筛,使用普通RT-PCR方法,分别用VP1、RdRp区和跨RdRp-VP1区引物对部分NoV阳性标本进行分型鉴定。应用SPSS20.0统计学软件进行数据处理,并使用生物信息学软件对NoV基因序列进行同源性、系统进化及重组性分析。结果经实时荧光RT-PCR检测,709份粪便标本中NoV阳性的标本为265份,阳性率为37.4%(265/709)。基于NoV RdRp和VP1区的测序分析,NoV阳性标本中共检出7种不同的基因型,分别为GⅡ.P16-GⅡ.2、GⅡ.P12-GⅡ.3、GⅡ.Pe-GⅡ.4_Sydney 2012、GⅡ.P7-GⅡ.6、GⅡ.P8-GⅡ.8、GⅡ.P21-GⅡ.13和GⅡ.P17-GⅡ.17型,其中以GⅡ.P16-GⅡ.2型所占比例最高,为30.6%(34/111),其次为GⅡ.Pe-GⅡ.4_Sydney 2012型(27.0%,30/111)和GⅡ.P12-GⅡ.3型(24.3%,27/111)。本研究发现2株新的NoV重组株GⅡ.P21-GⅡ.13,其重组位点位于ORF1/ORF2的交接区。NoV在每个月份皆有流行,但每个月份的主导基因型不尽相同。男女患儿的NoV阳性检出率差异无统计学意义(P=0.329),而不同年龄组患儿的NoV阳性检出率差异有统计学意义(P=0.011),以>11~12岁和>2~3岁年龄组所占比例最高。结论2018年10月至2019年9月,上海地区以GⅡ.P16-GⅡ.2、GⅡ.Pe-GⅡ.4_Sydney 2012和GⅡ.P12-GⅡ.3 NoV重组株流行为主,并发现2株新的NoV重组株GⅡ.P21-GⅡ.13散在流行。跨ORF1/ORF2的基因序列测定,有助于更好地了解NoV的基因分型和重组情况。Objective To investigate the genetic characteristics of norovirus(NoV)in children with acute gastroenteritis in Shanghai.Methods A total of 709 stool specimens were collected from outpatients with acute gastroenteritis in Children′s Hospital of Shanghai from October 2018 to September 2019.Real-time RT-PCR was used for qualitative detection of NoV,and RT-PCR was used to identify the genotypes of NoV with the primers of VP1 gene,RdRp region and RdRp-VP1 region.SPSS20.0 statistical software was used for data processing and bioinformatics software was used for homology,phylogenetic and recombination analysis of NoV gene sequences.Results NoV was detected in 265 out of the 709 stool specimens with a positive rate of 37.4%.Sequence analysis of RdRp region and VP1 gene showed that seven different genotypes including GⅡ.P16-GⅡ.2,GⅡ.P12-GⅡ.3,GⅡ.Pe-GⅡ.4_Sydney 2012,GⅡ.P7-GⅡ.6,GⅡ.P8-GⅡ.8,GⅡ.P21-GⅡ.13 and GⅡ.P17-GⅡ.17 were detected from 111 NoV-positive specimens.The predominated genotype was GⅡ.P16-GⅡ.2(30.6%,34/111),followed by GⅡ.Pe-GⅡ.4_Sydney 2012(27.0%,30/111)and GⅡ.P12-GⅡ.3(24.3%,27/111).Two new NoV recombinant strains belonging to GⅡ.P21-GⅡ.13 genotype were identified and the recombination site was in the junction region of ORF1 and ORF2.NoV infection occurred every month,but the predominant genotype was different.No significant difference in the positive rates of NoV was found between male and female patients(P=0.329).However,there were significant differences between different age groups(P=0.011)and the children in the age groups of>11-12 years old and>2-3 years old had higher rates of NoV infection.Conclusions The predominated recombinant NoV strains belonged to GⅡ.P16-GⅡ.2,GⅡ.Pe-GⅡ.4_Sydney 2012 and GⅡ.P12-GⅡ.3 genotypes,and two new recombinant NoV strains(GⅡ.P21-GⅡ.13)were found in Shanghai during October 2018 to September 2019.Gene sequencing across ORF1 and ORF2 was conducive to better understanding the NoV genotypes and recombination.

关 键 词：诺如病毒 急性胃肠炎 基因型 儿童 

分 类 号：R725.1[医药卫生—儿科]