传统细菌培养法、实时荧光PCR法在冷却水中的比较

Comparative Analysis of Legionella Pneumophila in Cooling Water Culture Method and Real-time PCR Method

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,P<0.05)。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。

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（ χ2=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.