传统细菌培养法实时荧光PCR法在冷却水中比较.docVIP

传统细菌培养法、实时荧光PCR法在冷却水中的比较 　　【摘要】 目的：冷却水中嗜肺军团菌传统细菌培养法和实时荧光PCR法检测结果比较。方法：选取某市范围的集中空调冷却水64份，同时对同一样品采用传统细菌培养法和实时荧光PCR法进行检测，用传统细菌培养法进行分离，对分离菌株再用实时荧光PCR法检测进行验证。结果：培养法嗜肺军团菌阳性率为12.50%（8/64），可疑阳性菌14株；进行实时荧光PCR法检测嗜肺军团菌阳性率为34.38%（22/64），两种检测方法比较差异有统计学意义（ 字2=9.389，P0.05）。将培养法的8株阳性菌及14株可疑阳性菌进行实时荧光PCR法检测，结果均为阳性。22株嗜肺军团菌血清型分布以LP1为主，占菌株总数的68.18%（15/22），其次是LP3占菌株总数的13.64%（3/22）。结论：培养法适用于冷却水中军团菌的检测，实时荧光PCR法则适用于对军团菌阳性菌株的辅助验证。 　　【关键词】 嗜肺军团菌； 细菌培养法； 实时荧光PCR 　　【Abstract】 Objective：To compare the results of cooling water Legionella pneumophila bacteria traditional culture and real-time PCR and fluorescence detection.Method：64 copies of a city-wide central air conditioning cooling water were selected，the bacteria were isolated and identified by conventional culture method，and then isolates validated by real-time PCR assay.Result：Legionella pneumophila culture positive rate was 12.50% （8/64），14 suspicious positive isolates；real-time PCR method of legionella pneumophila positive rate was 34.38% （22/64），the two detection methods were compared，the difference was statistically significant（ 字2=9.389，P0.05）.The conventional culture method of 8 positive isolates and 14 suspicious positive isolates were were identified by real-time PCR，the results were all positive for legionella pneumophila.In the separation of 22 legionella pneumophila，the serotype distribution was main in LP1，accounting for 68.18% of the total isolates （15/22），then the followed was LP3 accounting for 13.64% （3/22） of the total strain.Conclusion：The culture method applicable for the detection of legionella in cooling water，the real-time PCR method applicable to legionella-positive strains assisted verification. 　　【Key words】 Legionella pneumophila； Bacterial culture； Real-time PCR 　　军团菌广泛存在于各种天然和人工水环境中，其中嗜肺军团菌是引起非典型肺炎的重要病原菌，具有极强的传染性和较高的病死率，严重危害人类的健康和生命安全，已成为世界性的公共卫生问题；嗜肺军团菌在美国首次被发现，它主要是通过建筑物的水系统进行传播，感染该菌后发病速度快，死亡率高，我国自1982年在南京首次证实军团病以来，已有资料证实军团菌污染冷却水与军团菌病的暴发密切相关[1]。2003年卫生部《公共场所集中空调通风系统卫生规范》中明确规定在空调冷却水中不得检出军团菌[2]。冷却水中军团菌传统实验