葡萄球菌肠毒素基因分型PCR检测技术的研究  

Study on PCR Method for Differentiating Genetype of Entertoxin of S.aureus

作　　者：李琳[1] 黄金海[1] 赵耘[2] 王聪明[1] 董艳娇[1] 王静思[1] 刘莹[1] 

机构地区：[1]天津大学农业与生物工程学院,天津300072 [2]中国兽医药品监察所,北京100081

出　　处：《食品科学》2008年第7期340-344,共5页Food Science

基　　金：天津市应用基础及前沿技术研究计划项目(08JCZDJC22600)

摘　　要：通过生物信息学分析,设计了不同基因型肠毒素检测引物;应用含溶菌酶的碱裂解法提取葡萄球菌DNA及梯度PCR方法提高了肠毒素检出率和特异性。从国内28株葡萄球菌分离株中共检测出14种不同型的肠毒素基因(sea-see、tsst-1、seg-sei、sek-sel、sen-seo、seq-ser和seu),未检测到sej和sem基因。毒素基因携带率为16.67%,同时携带4种及以上毒素基因的菌株有11株,占39.29%,其中sek、seq和sea毒素基因在所研究菌株中分布最广,分别占到15.50%、14.30%和10.70%。结果表明不同葡萄球菌菌株间毒素型的关联度不同,sek与seq,seb和sed,sec、sei、sen和seu基因型呈现密切相关,各型肠毒素的分布还与菌株分离来源有着密切联系。所建立的PCR方法可用于金黄色葡萄球菌肠毒素基因分型和分布的研究。Various kinds of primers were designed to detect the genetypes of Staphylococcal enterotoxins（SEs） by bioinformatics analysis of enterotoxin genes. Lysozyme was added in the process of DNA extraction to enrich the DNA, encoding SEs. The detection rate and specificity were improved by anneal temperature gradient PCR method. 14 Kinds of enterotoxins were detected from the 28 isolates which indicate that their corresponding primers can effectively amplify the target genes while having high specificities. The three most common types of enterotoxins （sek, seq and sea ） in this research are approximately 15. 50%, 14.30% and 10.70% respectively. Whereas none of the isolates possesses the sem or sej genes. Different genotypes of SEs in stains have different degrees of correlation, and the genotypes of sek and seq, seb and sed, sec, sei, sen and seu display intimate positive correlations. Moreover, the results also suggested that the distribution of SEs has a close relationship with the source, which the Staphylococcus aureus is isolated from. The developed PCR method is useful in differentiating the genetypes and the distribution of S.aureus isolates.

关 键 词：金黄色葡萄球菌 肠毒素(SEs) PCR 检测 

分 类 号：R155.5[医药卫生—营养与食品卫生学]