军团菌快速检测方法的建立及其在国境口岸的应用

Establishment and application of rapid method for detecting Legionella

目的:建立适合口岸出入境大厅及其他公共设施中央空调水的分子生物学检测方法,以实现中央空调水的快速检测,提高检测速度及灵敏度。方法:根据军团菌核酸序列多重比对结果设计并合成实时荧光PCR的引物和探针,分别对军团菌16SrDNA和Mip基因相应片段进行扩增;建立快速、特异的军团菌分子快速检测新方法,并将该方法应用于对口岸中央空调水进行快速检测。结果:本研究以16SrDNA目标序列检测结果判断样本中是否存在军团菌,以Mip目标序列检测样本中是否存在嗜肺军团菌;本研究的方法快速、特异,能在2 h内完成对空调水样的检测。结论:本方法大大提高了军团菌的检测速度,提高了检测灵敏度,非常适合应用于中央空调水的快速检测。

Objective：To establish the approach of real-time fluorescence polymerase chain reaction for detecting Legionella from the condensate water of central air conditioner at the frontier port or other public building.Methods：According to multiple comparison of Legionella nucleotide sequence,PCR primers and probes were designed and synthesized to amplify corresponding gene segments of 16SrDNA and Mip for rapid detection of Legionella and Legionella pneumophila respectively in condensate water of central air conditioner at the frontier port.Results：The method reached 2.4 pg per reaction for 16SrDNA gene segment detection and 24 pg per reaction for Mip to the sensitivity of Legionella pneumophile,and could examine nucleic acid of Legionella accurately within two hours.Conclusion：This method can improve the speed and sensitivity of Legionella test,which was suitable for field rapid test of condensate water of central air conditioner.