赵云;林小娟;刘尧;纪峰;张丽;徐爱强;陶泽新;温红玲;

• 1:山东大学齐鲁医学院公共卫生学院微生物检验学系山东省“十三五”高等学校感染性疾病防控重点实验室
• 2:山东省疾病预防控制中心山东省传染病预防控制重点实验室

摘要(Abstract)：

人星状病毒（Human astrovirus,HAstV）是引起急性胃肠炎的主要致病病毒之一，为了解HAstV的型别分布和遗传变异特征，本研究于2018年在济南市按月采集12份生活污水样本，浓缩处理后提取RNA，进行HAstV部分ORF2编码区的PCR扩增，阳性产物进行下一代测序（Next generation sequencing, NGS）分析，探索HAstV各型别的动态变化和系统发生学特征。结果显示所有污水中均检出HAstV核酸，NGS分析显示所测读长分属于HAstV-1至HAstV-6共6个型别，其中HAstV-1型占比最大（27.4%），但不同月份的优势型别有所差别。系统发生学分析显示HAstV-1、HAstV-2、HAstV-5型山东株序列属于多个遗传分支，提示这些型别有多个传播链在当地循环。本研究描述了2018年济南市本地流行的人星状病毒基因型分布和序列特征，研究结果证明基于扩增子NGS的环境监测是探索人群中HAstV分子流行病学特征的有效手段。

关键词(KeyWords)： 环境监测;人星状病毒;下一代测序;系统发生

Abstract:

Keywords:

基金项目(Foundation): 山东省自然科学基金面上项目（项目号：ZR2020MH337）,题目：新型人星状病毒的分子流行病学研究;; 泰山学者青年专家计划项目（项目号：tsqn202103187）,题目：基于同一健康理念和下一代测序技术新型病毒的发现、溯源和进化动态研究;; 国家自然科学基金面上项目（项目号：81573209）,题目：环境污水中人类胃肠炎病毒的型别多样性、遗传进化规律及其与人群疾病发生的关系~~

作者(Authors): 赵云;林小娟;刘尧;纪峰;张丽;徐爱强;陶泽新;温红玲;

DOI: 10.13242/j.cnki.bingduxuebao.004211

参考文献(References)：

• [1]王永霞，段招军，李宇宁.人星状病毒研究进展[J/OL].病毒学报，2012, 28(04):482-487. DOI:10.13242/j.cnki.bingduxuebao.002308.
• [2] Marczinke B, Bloys A J, Brown T D, Willcocks M M,Carter M J, Brierley I. The human astrovirus RNAdependent RNA polymerase coding region is expressed by ribosomal frameshifting[J]. J Virol, 1994, 68(9):5588-5595.
• [3] Tao Z, Wang H, Zhang W, Xu A. Novel astrovirus types circulating in Shandong Province(Eastern China)during 2016:A clinical and environmental surveillance[J]. J Clin Virol, 2019, 116:69-73.
• [4] Guix S, Caballero S, Villena C, BartoloméR, Latorre C, Rabella N, SimóM, Bosch A, PintóR M.Molecular epidemiology of astrovirus infection in Barcelona, Spain[J]. J Clin Microbiol, 2002, 40(1):133-139.
• [5] Afrad M H, Karmakar P C, Das S K, Matthijnssens J,Ahmed F, Nahar S, Faruque A S, Rahman M Z,Rahman M, Azim T. Epidemiology and genetic diversity of human astrovirus infection among hospitalized patients with acute diarrhea in Bangladesh from 2010 to 2012[J]. J Clin Virol, 2013, 58(4):612-618.
• [6] Fumian T M, Leite J P, Rose T L, Prado T,Miagostovich M P. One year environmental surveillance of rotavirus specie A(RVA)genotypes in circulation after the introduction of the Rotarix?vaccine in Rio de Janeiro, Brazil[J]. Water Res, 2011, 45(17):5755-5763.
• [7] Tao Z, Wang H, Li Y, Xu A, Zhang Y, Song L,Yoshida H, Xu Q, Yang J, Zhang Y, Liu Y, Feng L,Xu W. Cocirculation of two transmission lineages of echovirus 6 in Jinan, China, as revealed by environmental surveillance and sequence analysis[J].Appl Environ Microbiol, 2011, 77(11):3786-3792.
• [8] Mustafa H, Palombo E A, Bishop R F. Epidemiology of astrovirus infection in young children hospitalized with acute gastroenteritis in Melbourne, Australia, over a period of four consecutive years, 1995 to 1998[J]. J Clin Microbiol, 2000, 38(3):1058-1062.
• [9] Guo L, Xu X, Song J, Wang W, Wang J, Hung T.Molecular characterization of astrovirus infection in children with diarrhea in Beijing, 2005-2007[J]. J Med Virol, 2010, 82(3):415-423.
• [10]Zhou N, Lin X, Wang S, Wang H, Bi Z, Wang P,Chen P, Tao Z, Song L, Song Y, Xu A. Molecular characterization of classic human astrovirus in eastern China, as revealed by environmental sewage surveillance[J]. J Appl Microbiol, 2016, 120(5):1436-1444.
• [11]Majumdar M, Martin J. Detection by direct next generation sequencing analysis of emerging enterovirus D68 and C109 strains in an environmental sample from Scotland[J/OL]. Front Microbiol, 2018, 9:1956.
• [12]Fumian T M, Fioretti J M, Lun J H, Dos Santos I A L, White P A, Miagostovich M P. Detection of norovirus epidemic genotypes in raw sewage using next generation sequencing[J]. Environ Int, 2019, 123:282-291.
• [13]Mancini P, Bonanno Ferraro G, Iaconelli M, Suffredini E, Valdazo-González B, Della Libera S, Divizia M, La Rosa G. Molecular characterization of human sapovirus in untreated sewage in Italy by amplicon-based Sanger and next-generation sequencing[J]. J Appl Microbiol,2019, 126(1):324-331.
• [14]Iaconelli M, Valdazo-González B, Equestre M,Ciccaglione A R, Marcantonio C, Della Libera S, La Rosa G. Molecular characterization of human adenoviruses in urban wastewaters using next generation and Sanger sequencing[J]. Water Res, 2017, 121:240-247.
• [15]Ulloa J C, Matiz A, Lareo L, Gutierrez M F.Molecular analysis of a 348 base-pair segment of open reading frame 2 of human astrovirus. A characterization of Colombian isolates[J]. In Silico Biol, 2005, 5(5-6):537.
• [16]Meleg E, Jakab F, Kocsis B, Bányai K, Melegh B,Szucs G. Human astroviruses in raw sewage samples in Hungary[J]. J Appl Microbiol, 2006, 101(5):1123-1129.
• [17]Lodder W J, de Roda Husman A M. Presence of noroviruses and other enteric viruses in sewage and surface waters in the Netherlands[J]. Appl Environ Microbiol, 2005, 71(3):1453-1461.
• [18]Iwai M, Hasegawa S, Obara M, Nakamura K,Horimoto E, Takizawa T, Kurata T, Sogen S, Shiraki K. Continuous presence of noroviruses and sapoviruses in raw sewage reflects infections among inhabitants of Toyama, Japan(2006 to 2008)[J]. Appl Environ Microbiol, 2009, 75(5):1264-1270.
• [19]Aw T G, Gin K Y. Environmental surveillance and molecular characterization of human enteric viruses in tropical urban wastewaters[J]. J Appl Microbiol, 2010,109(2):716-730.
• [20]Lizasoain A, Tort L F, García M, Gómez M M,Cristina J, Leite J P, Miagostovich M P, Victoria M,Colina R. Environmental assessment of classical human astrovirus in Uruguay[J]. Food Environ Virol, 2015, 7(2):142-148.
• [21]张文强，张华宁，韩扬，李心朋，刘晓林，傅忠燕，毕振旺，徐爱强，王海岩. 2017年山东省三个城市食源性腹泻中胃肠炎病毒的病原谱特征分析[J].中华预防医学杂志，2020(02):169-174.
• [22]Du H, Xiong P, Ji F, Lin X, Wang S, Zhang L, Liu Y, Song Y, Tao Z, Xu A. Genetic diversity and molecular epidemiological characterization of group A rotaviruses in raw sewage in Jinan by next generation sequencing[J/OL]. Infect Genet Evol, 2021, 91:104814.