贵阳市食源性金黄色葡萄球菌的单酶切扩增片段长度多态性基因分型  

Genotyping of foodborne Staphylococcus aureus in Guiyang by single-enzyme amplified fragment length polymorphism analysis

作　　者：吴正强[1] 万晴姣[1] 刘丽芳[1] 王玉倩[1] 翁庆北[1] 

机构地区：[1]贵州师范大学生命科学学院,贵州贵阳550001

出　　处：《环境与健康杂志》2015年第5期435-438,共4页Journal of Environment and Health

基　　金：贵阳市科学技术计划项目(2010筑科农合同字第1-社-51)

摘　　要：目的建立金黄色葡萄球菌的单酶切扩增片段长度多态性(SE-AFLP)基因分型方法,并对贵阳市食源性金黄色葡萄球菌菌株基因型进行分析。方法于2013年7—8月,采集贵阳市124份食品样品进行金黄色葡萄球菌的分离和鉴定,并利用PCR技术进一步验证。筛选HindⅢ单酶切AFLP扩增引物,对分离菌株进行SE-AFLP扩增分析,并根据扩增后的多态性条带进行聚类分析。结果共检出32株金黄色葡萄球菌,通过筛选扩增H-C引物可将其分为31个AFLP基因型。以Dice系数≥0.69作为分群依据,共分为A^E 5个群,其中A群为优势群,包括15个基因型,占总基因型48.39%,B群占16.13%,C群和D群各占12.90%,E群占9.68%。结论该方法能较好地对金黄色葡萄球菌进行分型,耗时短(1~2 d)且不需特殊的限制性内切酶和仪器设备。贵阳市食源性金黄色葡萄球菌具有丰富的遗传多样性。Objective Single-enzyme amplified fragment length polymorphism （SE-AFLP） markers were developed to analyze genotyping of foodborne Staphylococcus aureus in Guiyang. Methods Staphylococcus aureus was isolated and identified from 124 food samples collecting in Guiyang in July and August 2013. In addition,PCR was performed for further confirmation. After the AFLP primer of the restriction enzyme Hind Ⅲ was screened, the whole genomic DNA of the strain was analyzed by the SE-AFLP method. The amplification fragments were used for cluster analysis. Results A total of 32 strains of foodborne Staphylococcus aureus detected from 124 food samples were divided into 31 AFLP genotypes by SE-AFLP with H-C primer. Based on the DICE coefficient ≥0.69,these genotypes were classified into five groups：A to E. Group A comprised 15 genotypes and occupied 48.39% of total,and Group B,C,D and E occupied 16.13% ,12.90% ,12.90% and 9.68% of total, respectively. Conclusion Considering the procedural convenience and shorter time consumption, the modified AFLP protocol is a reliable tool for genotype analysis of Staphylococcus aureus. SE-AFLP molecular markers can indicate the significant polymorphism and genetic diversity within foodborne Staphylococcus aureus in Guiyang.

关 键 词：金黄色葡萄球菌 单酶切扩增片段长度多态性 基因分型 遗传多样性 

分 类 号：R117[医药卫生—公共卫生与预防医学]