上海市集中式中央空调冷却塔水嗜肺军团菌污染状况分析

目的了解上海市宾馆、大型商场、超市及写字楼中央空调系统嗜肺军团菌的污染状况。方法采集上海市100家公共场所集中式中央空调冷却塔水100份,经酸处理,以GVPC、BCYE、血平板进行菌落筛选,并应用生化反应鉴定,最后进行血清学分型。结果100份冷却塔水检出嗜肺军团菌的样本数为75份,检出率达75％（75／100）,其中宾馆、大型商场、超市、写字楼阳性率分别为80．6％、72．7％、72．7％、70％。75份检出水样中分离到76株军团菌,血清型别呈多样化,但以LPl为主,同一份水样同时检出2个型别的占总检出水样的1．3％（1／75）。结论上海市公共场所中央空调冷却水中嗜肺军团菌污染严重,血清型别呈多样化,对人群健康构成潜在威胁,需要引起重视,以防军团菌病的暴发流行。     

无锡市冷却塔水军团菌污染状况及菌型分析

目的了解无锡市不同场所冷却塔水中军团菌污染状况和血清型别。方法按照无锡市地理分布抽取32家公共场所和8家工厂作为监测点,共抽取冷却塔水529份,按照卫生部《集中空调通风系统卫生规范》附录A＂空调系统军团菌的检验分析方法＂进行检测。结果 529份冷却塔水军团菌的检出率达14.18%（75/529）,阳性率以商场和医院为高,分别为27.45%（14/51）和24.00%（18/75）。公共场所以7、8月份的阳性率为高,分别为18.75%（9/48）和25.25%（25/99）;工厂工艺性冷却塔水军团菌在全年各月份均有检出。军团菌血清型以LP1为主,占78.67%（59/75）。结论无锡市不同场所冷却塔水中存在军团菌污染,对人群健康构成潜在威胁,需引起重视。     

延安市286例呼吸科患者血清嗜肺军团菌抗体的检测

目的了解延安市嗜肺军团菌的感染情况。方法采用微量凝集法对286份呼吸科住院患者血清标本进行嗜肺军团菌Lp1型军团菌抗体测定。结果抗体阳性率为17．13％,其中男为18．56％,女为15．13％;在各年龄组均有嗜肺军团菌Lp1型抗体阳性者,其中40岁年龄组的感染率（26．09％）最高;呼吸科疾病中肺结核患者合并嗜肺军团菌Lp1感染率较高,为26．56％,各疾病类型之间感染率比较差异无统计学意义（P〉0．05）。结论延安市存在一定的嗜肺军团菌感染,需加强对军团病血清学的监测工作。     

应用单一和双重荧光定量PCR法快速检测军团菌

目的 建立单一和双重荧光定量PCR方法分别和同时进行军团菌属及嗜肺军团菌的检测.方法 利用军团菌属16 S rRNA基因和嗜肺军团菌mip基因设计引物和探针,两条基因探针分别标记FAM和HEX,并将相关反应体系和条件进行优化.分别应用单一基因探针(单一荧光定量PCR)和双重基因探针(双重荧光定量PCR)对嗜肺军团菌、非嗜肺军团菌及非军团菌进行检测,并验证两种方法的特异度、敏感度.应用双重荧光定量PCR检测空调水样滤膜样品和DNA提取样品,比较两者结果的一致性.结果 针对军团菌属及嗜肺军团菌,应用荧光定量PCR,16 S rRNA基因和mip基因均能较好的检出,16S rRNA和mip的最低检出限分别为8和10个拷贝.经优化得到了最佳反应体系.单一荧光定量PCR方法所检的8株嗜肺军用菌及4株非嗜肺军团菌16 S rRNA基因均为阳性,嗜肺军团菌mip基因阳性,非嗜肺军团菌mip基因阴性.双重荧光定量PCR方法所检的23株嗜肺军团菌中有2株为假阴性,9株非嗜肺军团菌和非军团菌属中有1株为假阳性.49份空调水样滤膜直接检测和提取DNA后检测的结果一致,其中26份水样军团菌阳性,20份为嗜肺军团菌,6份为非嗜肺军团菌;1份弗朗西斯菌检测HEX阳性(假阳性),占实际培养分离的1/26.结论 单一及双重荧光定量PCR法特异、快速、敏感,一次同时检测嗜肺与非嗜肺军团菌,满足对空调和环境水样军团菌监测的要求.     

应用单一和双重荧光定量PCR法快速检测军团菌

目的 建立单一和双重荧光定量PCR方法分别和同时进行军团菌属及嗜肺军团菌的检测.方法 利用军团菌属16 S rRNA基因和嗜肺军团菌mip基因设计引物和探针,两条基因探针分别标记FAM和HEX,并将相关反应体系和条件进行优化.分别应用单一基因探针(单一荧光定量PCR)和双重基因探针(双重荧光定量PCR)对嗜肺军团菌、非嗜肺军团菌及非军团菌进行检测,并验证两种方法的特异度、敏感度.应用双重荧光定量PCR检测空调水样滤膜样品和DNA提取样品,比较两者结果的一致性.结果 针对军团菌属及嗜肺军团菌,应用荧光定量PCR,16 S rRNA基因和mip基因均能较好的检出,16S rRNA和mip的最低检出限分别为8和10个拷贝.经优化得到了最佳反应体系.单一荧光定量PCR方法所检的8株嗜肺军用菌及4株非嗜肺军团菌16 S rRNA基因均为阳性,嗜肺军团菌mip基因阳性,非嗜肺军团菌mip基因阴性.双重荧光定量PCR方法所检的23株嗜肺军团菌中有2株为假阴性,9株非嗜肺军团菌和非军团菌属中有1株为假阳性.49份空调水样滤膜直接检测和提取DNA后检测的结果一致,其中26份水样军团菌阳性,20份为嗜肺军团菌,6份为非嗜肺军团菌;1份弗朗西斯菌检测HEX阳性(假阳性),占实际培养分离的1/26.结论 单一及双重荧光定量PCR法特异、快速、敏感,一次同时检测嗜肺与非嗜肺军团菌,满足对空调和环境水样军团菌监测的要求.     

嗜肺军团菌mip基因联合疫苗诱导的小鼠免疫应答

<正>军团病主要由嗜肺军团菌引起,嗜肺军团菌共有15个血清型,其中血清1型在人军团菌病中最常见,约占80%[1],可引起严重肺炎[2]。军团菌污染的水源通过加湿器、喷雾器、淋浴器、水龙头等载体形成气溶胶。人体吸入气溶胶后引起军团菌感染[3-4]。随着人们生活水平的提高,人工冷热水系     

南京地区肺科患者军团菌感染状况调查

了解南京市不同人群军团菌感染情况.选择南京市健康从业人员和肺科住院患者共计912份血清标本,采用微量凝集试验检测军团菌抗体效价,并对受检者进行流行病学调查.结果显示,南京市肺科患者军团菌抗体阳性率(46.3%,265/572)显著高于健康从业人员阳性率(39.4%,134/340)(P＜0.05).女性阳性率(45%)比男性阳性率(42.9%)稍高,但无显著性差异(P＞0.05).肺科患者中Lp 1型阳性率最高(14.2% ),其次为Lp 6型(8.9% ),Lp 10型最低(0.17%).健康从业人员中Lp 1型阳性率最高(12.4%),其次为Lp 3型(7.1%),Lp 6型(6.2%)和Lm型(5.0%)较低,Lp9未检出.结果提示,军团菌感染已经成为南京市肺科疾病重要的因素之一,并已对健康从业人员构成潜在危险.     

老年慢性支气管炎急性发作患者病原体检测结果分析

目的 分析引起老年慢性支气管炎急性发作的病原体,为临床提供诊断和治疗依据.方法 回顾2018年12月至2019年11月本院收治的199例老年慢性支气管炎急性发作患者的嗜肺军团菌1型(LP1)、肺炎支原体(MP)、Q热立克次体(COX)、肺炎衣原体(CP)、腺病毒(ADV)、呼吸道合胞病毒(RSV)、甲型流感病毒(IFA)、乙型流感病毒(IFB)、副流感病毒(PIV)抗体(IgM)水平以及痰病原菌培养结果,并对病原体耐药特点进行分析.结果 MP、LP1、PIV为最常见的非典型病原体,其IgM抗体阳性率分别为52.8％、26.1％和14.6％;2种以上(含2种)非典型病原体混合感染的检出率为30.7％,混合感染主要集中在MP及LP1合并病毒感染;非典型病原体抗体的阳性率受季节因素的影响,冬季的非典型病原体抗体的总阳性率显著高于其他季节(χ2=13.75,P<0.05),其中嗜肺军团菌的感染好发于冬季(χ2=15.03,P<0.05);副流感病毒的感染好发于冬季(χ2=10.67,P<0.05),其余非典型病原体抗体的阳性率未见明显季节性差异.细菌和非典型病原体混合感染的阳性率为6.5％(13/199);真菌和非典型病原体混合感染的阳性率为4.0％(8/199).结论 引起老年慢性支气管炎急性发作的病原体以MP、LP1、PIV等非典型病原体为主;非典型病原体的混合感染、非典型病原体与细菌的混合感染、非典型病原体与真菌的混合感染比较常见.     

石家庄市不同人群呼吸道感染病原体调查分析

目的 了解本地区急性呼吸道感染患者8种呼吸道病原体的流行情况.方法 选取2014年1月1日至2015年12月31日在石家庄市第一人民医院诊治的7545例呼吸道感染患者,对血清进行8种常见呼吸道病原体的IgM抗体检测.结果 本地区2014年检出率较高的呼吸道病原体依次是流感病毒B(17.5％)、流感病毒A(12.9％)、肺炎衣原体(11.8％),2015年检出率较高的呼吸道病原体依次是流感病毒B(20.6％)、肺炎支原体(20.0％)、流感病毒A(11.0％).2015年较2014年流感病毒A、流感病毒B、肺炎衣原体、嗜肺军团菌4种病原体IgM抗体阳性率有所降低,肺炎支原体IgM抗体阳性率有所升高;各年龄组八种病原体抗体阳性率均存在差异,与0～3岁组比较4～15岁组肺炎衣原体、流感病毒A、流感病毒B、副流感病毒、嗜肺军团菌、呼吸道合胞病毒IgM抗体阳性率较高,15岁以上组腺病毒、嗜肺军团菌、呼吸道合胞病毒IgM抗体阳性率较高;与4～15岁组比较15岁以上组腺病毒、嗜肺军团菌IgM抗体阳性率较高;且秋冬季与春夏季比较流感病毒A、流感病毒B、副流感病毒IgM抗体阳性率较高,而肺炎支原体IgM抗体阳性率较低.结论 本地区呼吸道感染的病原体以流感病毒A(INFA)、流感病毒B、肺炎支原体为主,不同年份、不同季节、不同年龄组病原体的种类和感染率具有一定差异性.     

实时荧光PCR快速检测嗜肺军团菌的研究

目的 建立TaqMan-MGB探针实时荧光PCR快速检测嗜肺军团菌技术,为临床和环境样品检测嗜肺军团菌提供可实用工具.方法 在对嗜肺军团菌mip序列进行分析、比较基础上,设计一对特异性引物和TaqMan-MGB探针,通过实时荧光PCR反应条件和反应体系的优化,实现对嗜肺军团菌的快速检测;用克隆到pMD-19T载体上的嗜肺军团菌mip基因阳参片段和不同菌株验证方法的敏感性和特异性.结果 当用热裂解法提取DNA,25μl的反应体系中包括上、下游引物(20μmol/L)各0.6μl,探针(20μmol/L)0.4μl,模板DNA 6.0μl,反应条件为预变95℃20 S,变性95℃10 s,退火50℃ 40 s,40个循环时,TaqMan-MGB探针实时荧光PCR技术对嗜肺军团菌mip基因阳参片段最低检测浓度为0.71拷贝/μl,其循环阈值(Ct值)与模板浓度具有极好的对应关系(r=0.999);1株嗜肺军团菌标准株、12株嗜肺军团菌分离株的Ct值在13.23～16.04之间,而包括金黄葡萄球菌、鼠伤寒沙门菌、副溶血性弧菌、大肠埃希菌、铜绿假单胞菌、痢疾志贺菌共计76株其他菌PCR Ct值均大于30;整个检测过程仅需1.5 h.结论 TaqMan-MGB探针的嗜肺军团菌实时荧光PCR检测方法具有特异性和敏感性、易操作、结果准确可靠等优点,可用于嗜肺军团菌检测.     

茂名市公共场所中央空调冷却水中军团菌污染状况调查

目的调查茂名市公共场所大型中央空调冷却水中军团菌的污染状况及主要血清型。方法于2006年8月至2007年7月．共采集茂名市区部分商场、酒店等中央空调冷却水24份，应用GVPC、BCYE、BCY培养基进行军团分离培养、生化鉴定及血清分型。结果冷却塔冷却水中军团菌的污染率达37．5％（9／24），血清型有LP1、U两种，以LJ（6／9）为主。结论茂名市公共场所大型中央空调冷却水中军团菌检出率较高，对市民健康构成了威胁。     

2005-2009年广州市星级宾馆二次供水卫生状况分析

目的了解广州市星级宾馆二次供水的卫生状况,为亚运期间制订针对性的卫生保障措施提供依据。方法收集广州市近5年来星级宾馆二次供水卫生监测的结果,整理数据并进行分析比较。结果5年来星级宾馆二次供水的总体合格率为86.05%,菌落总数的合格率最低为85.47%,其次为耗氧量和游离氯,第二、三季度的合格率明显低于第一、四季度。结论宾馆二次供水在贮存输送过程中受到的污染主要是微生物污染,应加强控制措施以保障中外宾客的健康。     

广州市市售食品食源性致病菌污染状况调查

目的了解广州市市售食品中食源性致病菌的污染状况。方法采用随机抽样方法分别对我市10个区以及两个地级市的集贸市场、超市、宾馆饭店及个体熟食销售点的7类食品共363份样品中沙门氏菌、金黄色葡萄球菌、副溶血性弧菌、单增李斯特菌、肠出血性大肠杆菌O157∶H7、空肠弯曲菌等6种致病菌依据国标的方法进行了监测。结果363件样品中检出致病菌72株。其中副溶血性弧菌检出率为33.93%,主要污染水产品;金黄色葡萄球菌为3.00%,主要污染熟食品;沙门氏菌和单增李斯特菌检出率分别为1.93%和0.66%;未检出大肠杆菌O157：H7和空肠弯曲菌。在各类食品中,致病菌带菌数有显著性差异（以χ^2检验,P〈0.005）。水产品致病菌带菌率最高,以副溶血性弧菌为主;其次为生肉类,以检出沙门氏菌为多。结论我市市售食品存在食源性致病菌污染,水产品和生肉类是主要污染食品。应加强市售食品监督管理,以减少可能引起食源性疾病的因素。     

Prevalence of Legionella pneumophila serogroup 1 in water distribution systems in Izmir province of Turkey

Legionella pneumophila serogroup 1 occurrence has been investigated in 168 hot water samples from 24 hotels, situated in 6 counties in Izmir province of Turkey, from 15 June to 30 September of the year 2000. Sampling was carried out at 15-day intervals and seven samples were taken from each of the hotels' hot water reservoirs and hot water networks. The samples were (1 L) concentrated using polycarbonate filters (mesh size 0.22 microm). Isolation was achieved using selective medium, GVPC agar. The samples were concentrated by membrane filtration, divided into three portions and cultured without pretreatment, after acid treatment, and after heat treatment, on GVPC agar. One hundred and ten isolates were identified as L.pneumophila sg 1 using the Legionella Latex Test (Oxoid). Arbitrarily primed PCR (AP PCR) was employed to assess the clonal relationship between Legionella pneumophila sg 1 isolates from the hot water samples of the hotels. Three genotypes of L. pneumophila sg 1 isolates were identified. With a high prevalence of type A, 22 hotels were found to be colonized with L. pneumophila serogroup 1, while only 2 were free from the bacteria.     

Comparative study of procedures for isolation and cultivation of Legionella pneumophila from tap water in hospitals.

For the isolation and cultivation of Legionella pneumophila from tap water in hospitals, we compared different media and selection techniques. A second part of the study compared the L. pneumophila yields from different water samples at identical sites. A total of 210 water samples (500 ml each) were collected from two selected sites in each of 21 hospitals. Warm water samples were collected after flow times of 0, 5, 10, and 15 min; in addition, one cold water sample was collected. Filtration was used to concentrate all samples. Following filtration, 0.1 and 1 ml each of untreated samples, heat-treated samples (3 min, 59 degrees C), and acid-treated samples (pH 2.2, 15 min) were spread onto the selective media MWY (SR 118; Oxoid) and BMPA alpha (SR 111; Oxoid), and samples from 12 hospitals were also spread onto GVPC medium (SR 152; Oxoid). A total of 72 (34%) of the 210 samples from 12 hospitals were positive. With respect to the positive Legionella cultures, there was no significant difference between the selective media MWY, BMPA alpha, and GVPC. With the BMPA alpha supplement, more samples were positive following heat treatment (P < 0.05) or acid treatment (P < 0.05) than without any further treatment. For the maximum yield of Legionella colonies with minimum additional microbial flora, acid treatment was the most effective, and by all methods, the GVPC supplement was the most selective. For routine water tests in hospitals for differentiating between systemic and local contamination, acid treatment of the concentrated samples, the use of different selective media, and the correct selection of sampling sites are recommended.     

2007年广州市公共场所用品用具卫生状况调查

目的了解2007年广州市各类公共场所用品用具的卫生状况。方法按照《公共场所卫生监测规范》和《公共场所卫生标准检验方法》的要求对公共场所用品用具进行抽样和检测，按照《公共场所卫生标准》进行分析和评价。结果2007年共监测广州76家公共场所单位的1430份用品用具，合格1326份，总合格率为92．73％。不同公共场所中，用品用具合格率最高为旅店业客房（95．38％），公共浴室和理发、美容室用品用具的合格率明显低于客房，差异有统计学意义（P〈0．05），娱乐场所用品用具合格率最低（仅为70％），与客房相比差异有统计学意义（P〈0．001）；不同种类用品用具中，杯具合格率最低（85．82％），但与其他各类相比差异无统计学意义。不合格检验项目主要是细菌总数，其次为大肠菌群。结论广州市公共场所用品用具卫生质量仍存在一些问题，应加强公共场所用品用具清洗、消毒卫生的监管工作。     

Molecular comparison of Legionella pneumophila serogroup 1 isolated in Tunisia

Legionella pneumophila is a common cause of hospital and community-acquired pneumonia, being transmitted by inhalation of aqueous aerosols. Most outbreaks are linked to contaminated hot water systems and cooling towers. Our study was about the molecular typing of 35 strains of L. pneumophila including four clinical isolates and 31 environmental strains isolated from the distribution systems of 14 hotels. Among the clinical strains, two have the same pattern, however, all were different from the studied environmental strains. For the 31 environmental strains, ten patterns were obtained. Among which, a same pulsotype was found for four strains isolated from four different establishments. In addition, two different pulsotypes were found for strains isolated from the same establishment. The pulsed-field gel electrophoresis showed the existence of various patterns. Although cases of legionellosis were declared in these hotels, there are no epidemiological links between the clinical and environmental strains.     

深圳市淡水鱼中香港海鸥菌污染状况调查

目的了解深圳市淡水鱼受香港海鸥菌污染的状况。方法于2011年2-3月共采集深圳市2家农贸批发市场和1家商场淡水鱼共计329份,将鱼肠中、后段内容物直接涂布改良麦康凯琼脂平板,凡生化符合的同时用特异性荧光PCR确诊香港海鸥菌。结果在329份淡水鱼中共检出香港海鸥菌7份,总阳性率为2.13%,其中塘虱鱼的阳性率最高为7.50%,鳙鱼的阳性率为2.50%,草鱼的阳性率为1.22%,其它淡水鱼未检出。7株香港海鸥菌生化反应阳性的,荧光PCR结果均为阳性。结论深圳市场淡水鱼中香港海鸥菌的污染不容忽视,应加强对淡水鱼产品中香港海鸥菌污染的监测,及时进行食品安全的预防和预警。     

Microbiological investigations on a nosocomial case of Legionella pneumophila pneumonia associated with water birth and review of neonatal cases

A case of Legionella pneumophila 1 pneumonia, confirmed by positive serology and urinary antigen, occurred in a 7-day old neonate after water birth in hospital. As respiratory samples were not available for culture, further microbiological investigations were performed in neonate and environment, in order to recognize the source of infection. The hospital water supply was contaminated by L. pneumophila 1 strains (300-2000 cfu/L) of two monoclonal subtypes of Pontiac subgroup. L. spiritensis (10-225 cfu/L) was isolated from cold tap water of the patient's home. PCR from tap and humidifiers water of the patient's home was positive for Legionella spp, but not for L. pneumophila. Because L. pneumophila 1, responsible of child infection, was only isolated from the hospital pool water for waterbirthing, we conclude that the infant acquired the nosocomial legionellosis by prolonged delivery in contaminated water, perhaps by aspiration. Infection control measures for waterbirthing are highly recommended. A review of neonatal case of legionellosis is also presented. As this rare infection may have a high fatality rate if unrecognized, pediatricians should be aware of the possibility of the legionellosis in newborns.     

广州市哨点医院腹泻儿童隐孢子虫感染的分子

目的了解广州某哨点医院门诊腹泻儿童隐孢子虫感染现状和分子流行病学特征。方法收集2012年6月至2013年5月广州市某哨点医院门诊腹泻儿童的人口资料学和粪便标本。采用巢氏聚合酶链反应（NestedPCR）方法检测粪便标本中的隐孢子虫卵囊。结果348份粪便标本中隐孢子虫的检出率为3．45％（12／348）,发病年龄以3岁以内为主（n=10）,检出率在患者的性别、年龄、月份间差异均无统计学意义,时间分布有2个高峰,分别为3—4月和11—12月。检出的12株隐孢子虫经测序分型,8株为C．hominis,4株为C．parvum,其中Ia亚型6株,Ib亚型2株,lIa亚型3株,IId亚型1株。结论广州某哨点医院腹泻儿童存在隐孢子虫感染,高发于春季和冬季,以3岁以内婴幼儿为主,主要流行C．hominis。