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目的建立多重PCR方法检测金黄色葡萄球菌α-溶血素、β-溶血素、h-溶血素基因以及16S rDNA,了解食源性金黄色葡萄球菌中3种溶血素基因的分布情况和溶血表型,并对菌株进行聚类分析。方法建立多重PCR方法检测148株食源性金黄色葡萄球菌的hla、hlb和hemolysin基因,并研究其分布情况,同时用血平板法检验其溶血表型,对数据进行汇总分析。结果148株菌中131株检出hla基因(88.51%),90株菌检出hlb基因(60.81%),28株菌检出hemolysin基因(18.92%);表型溶血的有131株(88.51%),不溶血的有17株(11.49%);以溶血素基因型和溶血表型为聚类因素,将148株菌以100%相似度为界分为A～L共12个型,其中以A型为主,有58株(39.19%),B型37株(25.00%),C型18株(12.16%)。结论该四重PCR方法特异性高,快速简便,能满足高通量菌株筛选的要求;食源性金黄色葡萄球菌普遍携带hla基因,聚类分析以A型和B型为主。
Objective To establish a multiplex PCR method for the detection of a-hemolysin, β-hemolysin, h-hemolysin gene and 16S rDNA of Staphylococcus aureus to understand the distribution of three hemolysin genes and the hemolysis phenotype in food-borne Staphylococcus aureus , And cluster analysis of strains. Methods Multiplex PCR was used to detect the hla, hlb and hemolysin genes of 148 strains of foodborne Staphylococcus aureus. The distribution of hla, hlb and hemolysin were studied. The hemolysin phenotypes were determined by the blood plate method, and the data were summarized and analyzed. Results Among the 148 strains, hla gene (88.51%) was detected in 131 strains, hlb gene (60.81%) in 90 strains, hemolysin gene (18.92%) in 28 strains, and 131 strains (88.51% ), And 17 (11.49%) did not hemolysis. Based on the hemolysin genotypes and hemolysis phenotypes, 148 strains were divided into 12 types of A ~ L by 100% similarity, among which There were 58 strains (39.19%), 37 strains (25.00%) of B type and 18 strains (12.16%) of C type. Conclusion The quadruple PCR method is highly specific, rapid and simple, and can meet the requirements of screening high-throughput strains. Food-borne Staphylococcus aureus generally carries hla gene. Cluster analysis mainly focuses on type A and type B.