金黄色葡萄球菌NorA蛋白免疫检测方法的建立

目的建立金黄色葡萄球菌耐药转运蛋白NorA免疫检测方法。方法利用琼脂稀释法测定诺氟沙星（NFLX）对临床分离的6株金葡菌和标准菌株ATCC25923的MIC。对norA基因进行克隆、表达，提取表达产物包涵体，并进行了纯化。将纯化的表达蛋白作为免疫原免疫家兔，获取NorA蛋白的抗体血清。利用制得的抗体通过Western blotting检测临床分离的6株金葡菌的NorA蛋白表达水平。结果氟喹诺酮类药物耐药水平较高的临床分离金葡菌菌株的NorA蛋白表达水平一般较高，敏感菌株的耐药水平较低且接近。结论免疫检测方法可用于检测金葡萄菌耐药转运蛋白NorA的表达水平。

Immunodetection of NorA protein in Staphylococcus aureus

Objective To establish a method of immunodetection of NorA protein in Staphylococcus aureus.Method Minimal inhibitory concentration(MIC) of norfloxacin against 6 clinical Staphylococcus aureus and control strain ATCC25923 was determined with disk diffusion and agar dilution susceptibility tests.The norA gene was cloned and expressed.The inclusion body was obtained,purified and used to immunized(rabbits).The antibody blood serum was detected by ELISA.Result The NorA antibody was obtained.Western blotting was used to detect the expression levels of NorA protein in 7 Staphylococcus aureus strains.The expression levels of NorA protein is higher in strongly resistant isolates,and the expression level of NorA protein is lower and almost the same as that in susceptible strains.Conclusion The immunodetection method can be used for the detection NorA protein in Staphylococcus aureus.