生活饮用水中嗜肺军团菌的酶底物测定方法研究
摘要：采用加标比对试验探索培养温度和时间对酶底物法检测饮用水中嗜肺军团菌的影响；在确定最适培养条件后,采集饮用水水样进行检测并通过16SrDNA序列GenBank比对试验对阳性结果进行确证。结果表明,酶底物法检测饮用水中嗜肺军团菌的最适培养条件为39℃恒温培养7d,嗜肺军团菌检出浓度与理论浓度基本一致,加标回收率范围为98.4%～108.8%；对北方城市部分饮用水水样进行嗜肺军团菌检测,检测发现阳性水样4份,经乳胶凝集血清分型试验16SrDNA序列分析确证均为嗜肺军团菌。
关键词：饮用水嗜肺军团菌酶底物法培养条件16SrDNA
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[2] NAKAMURA I,MIURA Y,UMEDA A,et al.The Legionella contamination of tap water in a brand-new hospital in Japan before patients move in[J].Infection Control and Hospital Epidemiology,2020,41(8):998-999．
[3] PASQUALINA,LAGAN,ALESSIO,et al.Environmental Surveillance of Legionellosis within an Italian University Hospital-Results of 15Years of Analysis[J].International Journal of Environmental Research and Public Health,2019,16(7):1103．
[4] GIROLAMINI L,MAZZOTTA M,LIZZADRO J,et al.Sit bath systems:A new source of Legionella infection[J].PLoSONE,2020,15(11):e0241756．
[5] DEEM S.Preparing for COVID's Effect on Legionella and Building Water Systems[J].Journal-American Water Works Association,2020,112(9):60-62．
[6] DEY R,ASHBOLT N J.Legionella Infection during and after the COVID-19Pandemic[J].2020．
[7] CUNHA B A,BURILLO A,BOUZA E.Legionnaires'disease[J].Lancet(London,England),2016,387(10016):376-385．
[8] YU V L,PLOUFFE J F,PASTORIS M C,et al.Distribution of Legionella species and serogroups isolated by culture in patients with sporadic community-acquired legionellosis:an international collaborative survey[J].The Journal of Infectious Diseases,2002,186(1):127-128．
[9] GB 5749-2006生活饮用水卫生标准[S]．中华人民共和国卫生部,中国国家标准化委员会．北京：中国标准出版社,2007．
[10] EPA 816-F-09-004.National primary drinking water regulations[S].Washington DC,2009．
[11] WHO.Guidelines for drinking-water quality[M].4th edition.Switzerland:WHO Press,2017．
[12] KIRSCHNER A K T.Determination of viable legionellae in engineered water systems:Do we find what we are looking for?[J].Water Research,2016,93:276-288．
[13] 吕恒,张锦海,陈凤娟,等．军团菌检测方法概述[J]．东南国防医药,2014(2):177-180．
[14] SCATURRO M,BUFFONI M,GIROLAMO A,et al.Performance of Legiolert Test vs.ISO 11731to Confirm Legionella pneumophila Contamination in Potable Water Samples[J].Pathogens,2020,9(9):690．
[15] KIRSTEN S,STEFAN P,BERND L,et al.Comparison of the Legiolert/Quanti-Tray MPN test for the enumeration of Legionella pneumophila from potable water samples with the German regulatory requirements methods ISO 11731-2and ISO 11731[J].International Journal of Hygiene and Environmental Health,2018,221(7):1047-1053．
[16] PETRISEK R,HALL J.Evaluation of a most probable number method for the enumeration of Legionella pneumophila from North American potable and nonpotable water samples[J].Water Health,2018,16(1):25-33．
[17] INOUE H,BABA M,TAYAMA S.Evaluation of Legiolert for Quantification of Legionella pneumophila from Bath Water Samples[J].Biocontrol Science,2020,25(3):179-182．
[18] MARK W.LeChevallier.Monitoring distribution systems for Legionella pneumophila using Legiolert[J].AWWA Water Science,2019,1(1)．
[19] LECHEVALLIER M W.Occurrence of culturable Legionella pneumophila in drinking water distribution systems[J].AW-WA Water Science,2019,1(3)．
[20] SARTORY D P,SPIES K,LANGE B,et al.Evaluation of a most probable number method for the enumeration of Legionella pneumophila from potable and related water samples[J].Lett Appl Microbiol,2017,64(4):271-275．
[21] Spies K,Pleischl S,Lange B,et al.Comparison of the Legiolert TM/Quanti-TrayMPN test for the enumeration of Legionella pneumophila from potable water samples with the German regulatory requirements methods ISO 11731-2and ISO 11731[J].Int J Hyg Environ Health,2018,221(7):1047-1053．
[22] MATHYS W,STANKE J,HARMUTH M,et al.Occurrence of Legionella in hot water systems of single-family residences in suburbs of two German cities with special reference to solar and district heating[J].Int J Hyg Environ Health,2008,211(1/2):179-185．
[23] BUSE H Y,ASHBOLT N J.Differential growth of Legionella pneumophila strains within a range of amoebae at various temperatures associated with in-premise plumbing[J].Letters in Applied Microbiology,2011,53(2):217-224.
Study on the determination method of enzyme substrate of Legionella pneumophilain drinking water
Abstract: In this study,we used a spiked comparison test to explore the effects of culture temperature and time on the enzyme substrate method for detecting Legionella pneumophila in drinking water;After determining the most suitable culture conditions,we collected drinking water samples for testing and confirmed the positive results by 16 SrDNA sequence GenBank comparison test.The results showed that the optimal culture condition for the detection of Legionella pneumophila in drinking water by the enzyme substrate method was 39℃constant temperature culture for 7 days,the detected concentration of Legionella pneumophilais basically the same as the theoretical concentration and the spiked recovery rate ranged from 98.4% to 108.8%;This study detected Legionella pneumophilain some drinking water samples from northern cities and found 4 positive water samples.Both methods of latex agglutination serotyping test and 16 SrDNA sequence analysis are confirmed that they were all Legionella pneumophila.
Keywords: Drinking water; Legionella pneumophila; Enzyme substrate; Culture conditions; 16SrDNA;
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