酶底物法定量检测嗜肺军团菌的应用研究

Applied research on enzyme substrate method in quantitative detection of Legionella pneumophila

目的开展嗜肺军团菌检测新方法的应用研究,利用酶底物法定量检测技术应用于辖区公共场所不同水样的嗜肺军团菌检测中。方法 2018年—2021年收集辖区公共场所淋浴热水、水龙头水样以及集中空调冷凝水、冷却水样,采用酶底物法进行检测,对检出的阳性样本经细菌分离技术与TaqMan荧光定量PCR实验验证确认。结果应用酶底物定量检测法在336件水样中检出阳性样本81件,阳性率为24.1%。嗜肺军团菌计数结果在<1 MPN/100 ml~2 272.6 MPN/100 ml。对81件阳性样本经细菌分离培养、血清分型实验、荧光定量PCR实验加以验证确认,无假阳性,方法特异度为100%。结论通过应用酶底物法检测嗜肺军团菌,操作简便、结果快速、定量、灵敏,具有推广应用的价值。

Objective To study the application of a new method for the detection ofLegionella pneumophila, and to apply the enzyme substrate method in the quantitative detection ofLegionella pneumophilain different water samples in public places under the jurisdiction.Methods From 2018 to 2021, water samples of shower hot water and faucet in public places, condensed water and cooling water in central air conditioners were collected and detected by enzyme substrate method, and positive samples were verified and confirmed by bacterial separation technology and TaqMan fluorescence quantitative PCR experiment.Results 81 of 336 water samples were detected by enzyme substrate quantitative detection method, with a positive rate of24. 1%. The count ofLegionella pneumophilaranged from < 1 MPN/100 ml-2 272. 6 MPN/100 ml. The 81 positive samples were verified and confirmed by bacterial isolation and culture, serum typing test and fluorescence quantitative PCR test, and no false positive was found. The specificity of the method was 100%.Conclusion Using enzyme substrate method to detectLegionella pneumophilais simple, rapid, quantitative and sensitive, which has the value of popularization and application.