摘要
目的建立嗜肺军团菌特异、快速检测方法,探讨南京地区公共场所环境水样嗜肺军团菌污染状况。方法根据嗜肺军团菌的mip基因保守序列设计引物和探针,通过对嗜肺军团菌标准株ATCC33152、嗜肺军团菌南京分离株及大肠埃希菌ATCC25922、鼠伤寒沙门菌ATCC14028、福氏2a志贺菌ATCC12022进行检测,验证该方法的特异性。并应用该法对南京地区某商场、宾馆、办公楼、医院共计13家的中央空调系统的冷却水、淋浴水和景观水共计88份环境水样进行了PCR检测及军团菌的分离培养。结果该法对嗜肺军团菌有特异性扩增曲线,而对其他细菌无阳性扩增曲线。88份水样中荧光定量PCR方法检测出28份阳性,阳性率为31.8%;传统培养方法分离出17株军团菌,分离率为19.3%。结论本文建立的荧光定量PCR检测方法具有良好的特异性,可用于环境水中军团菌污染状况调查。南京地区中央空调冷却水中检出率最高,存在发生军团菌病的可能,应引起足够重视。
Objective To develop a specific and rapid fluorescence quantitative real = time PCR assay in the detection of Leg/onella pneumophila, and to investigate the contamination of Legionella pneumophila in public places in Nanjing. Methods According to the conservative sequence of mip genes, the primers and the fluomgenic probe were designed and synthesized, and then the detections of Legionella pneumophila standard strain ATCC33152, Legionella pneumophila Nanjing isolates and Escherichia coli ATCC25922, Salmonella typhimurium ATCC14028, Shigella flexneri ATCC12022 （2a） were used to verify the specificity of the fluorescence quantitative real -time PCR assay. Then the method was used to test 88 environmental water samples collected from central air conditioning cooling water, shower water and landscape water in 13 shopping mails, hotels, office buildings and hospitals of Nanjing area. Results The results showed that there was a positive amplification curve for Legionella pneumophila standard bacteria and local isolates, but no positive amplification curve for other bacteria was observed. Of 88 samples, 28 positive samples were detected by fluorescence quantitative PCR method, with the detective rate of 31.8% ; and 17 isolates were cultivated by the traditional culture method, with the isolation rate of 19.3%. Conclusion The fluorescent quantitative PCR detection method established in this paper has good specificity and can be used for the investigation of Legionella pneumophila contamination in environmental water, and the detection rate of Legionella pneumophila was the highest in air conditioning cooling water. There is a possibility of Legionnaires disease, which should be paid enough attention t0.
作者
叶艳华
江晓
金萍
何敏
丁洁
张洪英
YE Yanhua;JIANG Xiao;JIN Ping;HE Min;DING Jie;ZHANG Hong ying(Nanjing Municipal Center for Disease Control and Prevention, Nanjing, Jiangsu 210003, China)
出处
《中国卫生检验杂志》
CAS
2018年第8期901-903,共3页
Chinese Journal of Health Laboratory Technology
基金
南京市医学科技发展课题(YKK11053)
关键词
嗜肺军团菌
荧光定量PCR
MIP基因
LegioneUa pneunugphila
Fluorescence quantitative PCR
mip gene
