16S rRNA基因测序分析技术在食品污染菌监测中的应用  

Application of 16S rRNA gene sequencing and sequence analysis in foodborne pathogens monitoring

作　　者：胡鹏威[1] 刘楚云[1] 何娇明 张祥 袁月明[1] 袁梦[1] 段永翔[1] HU Peng-wei;LIU Chu-yun;HE Jiao-ming;ZHANG Xiang;YUAN Yue-ming;YUAN Meng;DUAN Yong-xiang(Department of Microbiological Laboratory,Shenzhen Nanshan Center for Disease Control and Prevention,Shenzhen,Guangdong 518054,China)

机构地区：[1]深圳市南山区疾病预防控制中心微生物检验科

出　　处：《现代预防医学》2019年第11期2028-2031,2101,共5页Modern Preventive Medicine

基　　金：深圳市"医疗卫生三名工程"细菌感染性疾病检测及监测项目(SZSM201803081)

摘　　要：目的将16S rRNA基因测序分析技术应用于食品安全风险监测中非目标(相似、少见或非典型)致病菌的鉴定,为食源性疾病防控提供更多病原学证据。方法采集18份畜类、18份禽类和9份鱼类共45份生鲜样品,目标致病菌按照GB4789标准检测,非目标菌进行16S rRNA基因测序,将全长序列在GenBank中比对确定细菌种属,通过MEGA6构建细菌16S rRNA基因进化树,判定细菌种属间的亲缘关系。结果 45份生鲜样品中检出19株目标致病菌,包括沙门氏菌、空肠弯曲菌、单增李斯特菌和副溶血弧菌,而34株非目标菌16S rRNA基因全长序列能分别比对出13种细菌,其中8种19株菌属于致病或条件致病菌,包括霍乱弧菌、溶藻弧菌、结肠弯曲菌、奇异变形杆菌、粪肠球菌和铜绿假单胞菌,以及在深圳乃至省内首次报道的布氏弓形菌和海藻希瓦氏菌。结论深圳市售生鲜食品受到多种食源性致病菌污染,特别是非目标检出的致病菌,对此认为16S rRNA基因测序分析技术是一套有效的细菌监测适宜技术,能提高食源性疾病和食物中毒的防控水平。Objective To use the 16 S rRNA gene sequencing and sequence analysis in the identification of non-target foodborne pathogens in the food safety risk monitoring,so as to provide more etiological evidence for the prevention and control of foodborne diseases.Methods A total of 45 fresh foods were sampled,including the samples of 18 livestock,18 poultry and 9 freshwater fish.The target pathogens were detected according to the GB4789 standard.The non-target pathogens were confirmed by the PCR amplification and gene sequencing with specific 16 S rRNA universal primers.The sequencing data were aligned and compared in the GenBank database,so as to determine bacterial species.The 16 s rRNA trees of phyletic classification were established via the MEGA6 software,in order to reveal the genetic relationships of germs.Results Nineteen target pathogenic strains contained Salmonella,Campylobacter jejuni,Listeria monocytogenes and Vibrio parahaemolyticus were detected.A total of 34 non-target strains were classified into 13 species based on the whole 16 S rRNA sequences.Eight kinds of 19 non-target strains belonged to(opportunistic)pathogens,including Vibrio cholera,Vibrio alginolyticus,Campylobacter coli,Proteus mirabilis,Enterococcus faecalis and Pseudomonas aeruginosa,as well as the first reported Arcobacter butzleri and Shewanella algae in Shenzhen and even Guangdong.Conclusion The fresh foods of livestock,poultry and fish are contaminated by a wide variety of foodborne pathogens,particularly the non-target germs.Thus the 16 S rRNA gene sequencing and sequence analysis is considered as a set of appropriate technology for bacteria monitoring,which can enhance the prevention and control level of foodborne illnesses and outbreaks.

关 键 词：食品监测 食源性致病菌 16S RRNA 基因测序 分子鉴定 

分 类 号：R117[医药卫生—公共卫生与预防医学]