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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,Simpon相关系数为0.916 4。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 melting curve and apply it to the rapid detection of Staphylococcus aureus food poisoning. Methods According to the literature, the corresponding primers of VNTR of Staphylococcus aureus were synthesized and used to detect Staphylococcus aureus of known PFGE type. Four high-resolution sites were selected. On the basis of this, a molecular typing system of melting curve was established to detect 59 Staphylococcus aureus isolated from food poisoning. Results The molecular typing system of the melting curve had good precision. The standard deviation (s) of batch repeat experiment was 0.03 ~ 0.05 ℃, and s was 0.04 ~ 0.06 ℃. The daytime repeatability experiment s is 0.04 to 0.06 ° C. Meanwhile, 59 strains of Staphylococcus aureus could be divided into 19 types, including 11 popular clones and 8 sporadic clones. The Simpon correlation coefficient was 0.916 4. Conclusion The established system is rapid, sensitive and reproducible. It can be applied to the rapid tracing and screening of Staphylococcus aureus food poisoning.