高佳迎, 袁灵月, 陈艳新, 陈涌, 罗嘉远, 陈明亮, 陈敏. 2019—2020年上海市医院人工水环境中嗜肺军团菌的污染状况与分子分型研究[J]. 环境与职业医学, 2023, 40(7): 823-829. DOI: 10.11836/JEOM22515
引用本文: 高佳迎, 袁灵月, 陈艳新, 陈涌, 罗嘉远, 陈明亮, 陈敏. 2019—2020年上海市医院人工水环境中嗜肺军团菌的污染状况与分子分型研究[J]. 环境与职业医学, 2023, 40(7): 823-829. DOI: 10.11836/JEOM22515
GAO Jiaying, YUAN Lingyue, CHEN Yanxin, CHEN Yong, LUO Jiayuan, CHEN Mingliang, CHEN Min. Contamination status and molecular typing of Legionella pneumophila in artificial water environment in Shanghai hospitals from 2019 to 2020[J]. Journal of Environmental and Occupational Medicine, 2023, 40(7): 823-829. DOI: 10.11836/JEOM22515
Citation: GAO Jiaying, YUAN Lingyue, CHEN Yanxin, CHEN Yong, LUO Jiayuan, CHEN Mingliang, CHEN Min. Contamination status and molecular typing of Legionella pneumophila in artificial water environment in Shanghai hospitals from 2019 to 2020[J]. Journal of Environmental and Occupational Medicine, 2023, 40(7): 823-829. DOI: 10.11836/JEOM22515

2019—2020年上海市医院人工水环境中嗜肺军团菌的污染状况与分子分型研究

Contamination status and molecular typing of Legionella pneumophila in artificial water environment in Shanghai hospitals from 2019 to 2020

  • 摘要: 背景

    全球军团菌病发病率升高,人工水环境成为其暴发常见来源之一,而分子分型技术有助于预防和控制军团菌。

    目的

    了解上海市医院人工水环境中嗜肺军团菌的分子流行病学特征,为军团菌病的防控提供科学依据。

    方法

    于2019—2020年每年5—10月采集上海市14家医院人工水环境中水样,包括8家三甲医院,6家非三甲医院。共采集水样984份,包括312份冷却水、72份冷冻水和600份自来水。对水样进行嗜肺军团菌的分离培养和血清型鉴定并保藏,以嗜肺军团菌检出率作为污染状况指标。对保藏菌株进行复苏,得到81株存活菌株进行脉冲场凝胶电泳(PFGE)分型分析。

    结果

    共检出嗜肺军团菌水样124份,样本检出率12.60%,其中三甲医院检出率(16.54%,87/526)高于非三甲医院(8.08%,37/458)(χ2=15.91,P<0.001)。不同类型水样之间冷却水的检出率(23.40%)最高,检出率的差异具有统计学意义(χ2=61.19,P<0.001)。医院不同场所自来水检出率差异具有统计学意义(χ2=11.37,P<0.05)。2019年的检出率(15.06%,75/498)高于2020年(9.84%,49/498)(χ2=6.23,P<0.05)。5—10月中,8月的年平均检出率(16.46%)最高,10月的年平均检出率(8.54%)最低,不同月份之间检出率的差异无统计学意义(χ2=5.39,P=0.37)。不同区之间的检出率差异具有统计学意义(χ2=24.88,P<0.001)。共分离得到131株嗜肺军团菌,以血清型1型(80.15%,105/131)为主。对复苏存活的81株嗜肺军团菌进行PFGE分型,菌株条带相似性系数为41.30%~100%,分为29种PFGE带型(S1~S29),每种带型包括1~10株菌,S28为优势带型。聚类得到4个聚类簇(Ⅰ~Ⅳ),占所有菌株的66.67%(54/81),包含13个带型。

    结论

    上海市2019—2020年医院人工水环境中存在嗜肺军团菌持续污染,自来水中污染值得引起重视。检出嗜肺军团菌血清型以1型为主,PFGE分型揭示其存在基因多态性。因此,需加强医院人工水环境中嗜肺军团菌的监测及控制。

     

    Abstract: Background

    The incidence of Legionnaires' disease is increasing globally and artificial water environment is becoming a common source of outbreaks. Molecular typing techniques can help prevent and control Legionella.

    Objective

    To understand the molecular epidemiological characteristics of Legionella pneumophila in artificial water environment of Shanghai hospitals, and provide a scientific basis for the prevention and control of Legionnaires' disease.

    Methods

    Water samples were collected from artificial water environment in 14 hospitals from May to October each year from 2019 to 2020 in Shanghai. A total of 984 water samples were collected from 8 Grade-A tertiary hospitals and 6 non-Grade-A tertiary hospitals, including 312 samples of cooling water, 72 samples of chilled water, and 600 samples of tap water. The water samples were isolated and serotyped for Legionella pneumophila and preserved, and the positive rate of Legionella pneumophila in the samples was used as an indicator of contamination. The preserved strains were resuscitated and 81 surviving strains were obtained for pulsed field gel electrophoresis (PFGE) typing analysis.

    Results

    A total of 124 Legionella pneumophila positive water samples were detected, with a positive rate of 12.60%. The positive rate was higher in the Grade-A tertiary hospitals (16.54%, 87/526) than in the non-Grade-A tertiary hospitals (8.08%, 37/458) (χ2=15.91, P<0.001). The positive rate of cooling water (23.40%) was the highest among different types of water samples, and the difference was statistically significant (χ2=61.19, P<0.001). The difference in positive rate of tap water was statistically significant among different hospital departments (χ2=11.37, P<0.05). The positive rate in 2019 (15.06%) was higher than that in 2020 (9.84%) (χ2=6.23, P<0.05). From May to October, August had the highest annual average positive rate (16.46%) and October had the lowest (8.54%), but the difference in positive rates among months was not statistically significant (χ2=5.39, P=0.37). The difference in positive rate among districts was statistically significant (χ2=24.88, P<0.001). A total of 131 strains of Legionella pneumophila were isolated, with serotype 1 (80.15%, 105/131) predominating. Among the 81 surviving strains of Legionella pneumophila subjected to PFGE typing, the band-based similarity coefficients ranged from 41.30% to 100%. Among the 29 PFGE band types (S1-S29) recorded, each band type included 1-10 strains, and S28 was the dominant band type. Four clusters (I-IV) of PFGE band types were identified, accounting for 66.67% (54/81) of all strains and containing 13 band types.

    Conclusion

    Legionella pneumophila contamination is present in the artificial water environment of hospitals in Shanghai from 2019 to 2020, and the contamination in tap water deserves attention. The detected serotype of Legionella pneumophila is predominantly type 1, and PFGE typing reveals the presence of genetic polymorphism. Therefore, the monitoring and control of Legionella pneumophila in hospital artificial water environment should be strengthened.

     

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