金黄色葡萄球菌熔解曲线分子分型体系的建立及应用

Establishment and application of a molecular typing system of Staphylococcus aureus based on resolution melting

目的：建立金黄色葡萄球菌熔解曲线分子分型体系，应用于金黄色葡萄球菌食物中毒快速溯源。方法根据文献合成金黄色葡萄球菌 VNTR 位点的相应引物，用于检测已知 PFGE 型别的金黄色葡萄球菌，从中筛选出4个分辨力较高的位点。在此基础上建立熔解曲线分子分型体系，检测59株分离自食物中毒的金黄色葡萄球菌。结果熔解曲线分子分型体系具有良好的精密度，其批内重复性实验的标准差（s）为0．03～0．05℃，批间重复性实验的 s 为0．04～0．06℃，日间重复性实验的 s 为0．04～0．06℃。同时，熔解曲线分子分型体系能将59株金黄色葡萄球菌分为19个型别，包括11个流行克隆和8个散发克隆，Sim-pon 相关系数为0．9164。结论建立的体系快速、灵敏度高、重复性好，能应用于金黄色葡萄球菌食物中毒的快速溯源和筛查。

Objective To establish a molecular typing system of Staphylococcus aureus by using resolution melting for food-poi-soning fast tracing.Methods Primers were designed and synthesized according to the literature of VNTR in Staphylococcus au-reus ,and were used to perform molecular typing on the strains which had detected by PFGE,then 4 types of VNTRs were with higher discriminatory power were selected.On this basis,we established a molecular typing system for the detection of 59 Staphy-lococcus aureus isolated from food poisoning.Results The molecular typing system has good precision for detection.The standard deviation（s）of within-batch repetitive experiments were 0.03 -0.05 ℃,between-batch repetitive experiments were 0.04 -0.06℃,between-day repetitive experiments were 0.04-0.06 ℃.At the same time,the 59 strains of Staphylococcus aureus were divided into 19 types which were 11 epidemic clones and 8 sporadic clones.The correlation coefficient of Simpson was 0.916 4.Conclusion The molecular typing system for Staphylococcus aureus based on resolution melting was simple,fast and repeatable.It can be ap-plied to fast tracing and screen of Staphylococcus aureus in food poisoning.