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目的建立嗜肺军团菌Lp1型实时荧光定量PCR检测方法,以实现对中央空调冷却水中嗜肺军团菌Lp1型的快速检测。方法基于Lp1型嗜肺军团菌基因组ORF11区域设计特异性的引物和探针,建立体系,验证方法的灵敏度、特异性和重复性,并对模拟的中央空调冷却水水样进行检测。结果该方法的检测灵敏度达3.8×10 cfu/ml,具有良好的特异性,仅嗜肺军团菌Lp1型菌株呈阳性结果,重复检测平均Ct值变异系数。对模拟的水样进行检测,与方法的灵敏度相一致,且具有良好的重复性。结论该方法可实现对嗜肺军团菌Lp1型的快速检测。
Objective To establish a Lp1 real-time PCR method for detection of Legionella pneumophila in order to achieve rapid detection of L. pneumophila Lp1 in central air-conditioning cooling water. Methods Based on ORF11 region of Legionella pneumophila Lp1, primers and probes were designed to establish the system to verify the sensitivity, specificity and repeatability of the method. The simulated water samples of central air conditioning cooling water were also tested. Results The detection sensitivity of this method was 3.8 × 10 cfu / ml with good specificity. Only Legionella pneumophila Lp1 showed positive results, and the average coefficient of variation of Ct was detected repeatedly. The simulation of the water samples were tested, consistent with the sensitivity of the method, and has good repeatability. Conclusion This method allows rapid detection of Legionella pneumophila Lp1.