可视化金黄色葡萄球菌LAMP检测方法的建立及应用

发布时间：2018-01-19 11:01

本文关键词： 环介导等温扩增金黄色葡萄球菌耐热核酸酶　出处：《现代预防医学》2017年22期 　论文类型：期刊论文

【摘要】：目的建立一种基于颜色判断结果的金黄色葡萄球菌(简称金葡菌)快速、敏感和特异的环介导等温扩增(Loop-mediated isothermal amplification,LAMP)方法并应用于食源性疾病样品的金葡菌筛查。方法针对金葡菌耐热核酸酶(thermostable nuclease,nuc)基因特异性序列的6个区域设计2对引物,建立LAMP方法。利用LAMP和普通PCR方法同时检测金葡菌和对照组菌株来验证LAMP方法的特异性;将金葡菌菌液作10倍系列稀释后利用LAMP和PCR方法同时进行检测来评价两者的敏感度,利用LAMP和PCR方法分别对330份食源性疾病样品进行检测来比较两者的检出率。结果 LAMP方法可以利用肉眼进行结果判断,其特异性与普通PCR相当,但其敏感性比普通PCR高10倍,LAMP方法检测下限为10 cfu/ml,PCR方法检测下限为100 cfu/ml;LAMP和普通PCR方法对食源性疾病样品的检出率相符。结论可视化金葡菌LAMP方法是一个快速、敏感和特异的方法,可应用于食源性疾病样品中金葡菌的筛查。
[Abstract]:Objective to establish a rapid method for staphylococcus aureus (Staphylococcus aureus) based on color judgment. Loop-mediated isothermal amplification was amplified by sensitive and specific loop mediated isothermal amplification. Methods the method was used to screen Staphylococcus aureus from foodborne disease samples. Methods Thermostable nuclease was used against thermostable nuclease of Staphylococcus aureus. Two pairs of primers were designed for 6 regions of nuclear gene specific sequence. The LAMP method was established. The specificity of the LAMP method was verified by both LAMP and PCR methods for the simultaneous detection of Staphylococcus aureus and control strains. The sensitivity of Staphylococcus aureus solution was evaluated by LAMP and PCR methods after dilution of 10-fold series. LAMP and PCR methods were used to detect 330 foodborne disease samples to compare the detection rate of the two methods. Results LAMP method can be used to judge the results with the naked eye. Its specificity was similar to that of normal PCR, but its sensitivity was 10 times higher than that of PCR. The detection limit of lamp method was 10 cfu/ml. The detection limit of PCR method is 100cfur / ml; The detection rates of LAMP and PCR were consistent with those of foodborne disease samples. Conclusion the visual LAMP method is a rapid, sensitive and specific method for the detection of foodborne diseases. It can be used in the screening of Staphylococcus aureus in foodborne disease samples.
【作者单位】：长沙县食品安全检测中心;长沙市疾病预防控制中心;
【基金】：湖南省卫生计生委2015年度科研计划项目(No.B2015-153)
【分类号】：R155.5
【正文快照】： 金黄色葡萄球菌(简称金葡菌)是一种革兰阳性球菌,不但能够造成感染部位的化脓性炎症,还可以产生毒素引起食源性疾病[1]。金葡菌及其肠毒素引起的食源性疾病是重要的公共卫生问题,国家卫生和计生委通报情况显示,2015年全国共报告食物中毒突发公共卫生事件169起,中毒5 962人,死

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