实验动物金黄色葡萄球菌PCR检测方法的建立和应用

Establishment and application of PCR detection method for Staphylococcus aureus in laboratory animals

为了快速检测出实验动物临床样品中的金黄色葡萄球菌(SA),建立了一种普通PCR方法。同时合成3对引物,用金黄色葡萄球菌的阳性核酸DNA做模板,进行引物扩增,筛选出一对能扩增出270 bp单一目的片段的引物。逐一对该引物灵敏度、重复性和特异性进行了测试。实验结果显示,该方法具有高度的特异性,除了识别金黄色葡萄球菌基因组外,对小鼠其他常见病原菌均不发生交叉反应,具有优良的敏感性和稳定性。两个不同操作人员,在两个不同时间段,利用两台不同品牌的PCR仪器,用金黄色葡萄球菌阳性样品进行测试,结果显示此方法再现性良好。用该方法检测待测样本,效果也良好。

In order to quickly detect Staphylococcus aureus(SA)in clinical samples of laboratory animals,a common PCR method was established.Three pairs of primers were synthesized simultaneously,and the positive nucleic acid DNA of Staphylococcus aureus was used as a template for primer amplification.A pair of primers capable of amplifying a single target fragment of 270 bp was selected.The sensitivity,repeatability,and specificity of the primers were tested one by one.The experimental results show that this method has high specificity.Apart from identifying the genome of Staphylococcus aureus,it does not cross react with other common pathogenic bacteria in mice,and has excellent sensitivity and stability.Two different operators tested positive samples of Staphylococcus aureus using two PCR instruments of different brands at two different time periods,and the results showed good reproducibility of this method.The method is also effective in detecting the samples to be tested.