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。     

广州市部分公共场所集中空调通风系统卫生现况调查

目的了解广州市部分公共场所集中空调通风系统的卫生现况，为规范公共场所集中空调通风系统的卫生管理提供依据。方法随机抽取广州市部分大型公共场所的集中空调通风系统，对新风量进行检测；对送风中可吸入颗粒物（PM10）、细菌总数、真菌总数、β-溶血性链球菌进行检测并对总挥发性有机化合物（TVOC）、军团菌进行检测。结果部分公共场所集中空调通风系统新风量合格率为99．01％，送风中PM10合格率为95．53％．细菌总数合格率为92．64％，真菌总数合格率为90．70％，β-溶血性链球菌合格率为100％；空气净化消毒装置的TVOC合格率为81．05％；冷却水冷凝水中军团菌合格率为89．58％。结论公共场所集中空调通风系统应加强卫生管理。     

广州市亚运接待宾馆酒店淋浴热水军团菌污染状况分析

目的对广州市亚运接待宾馆酒店淋浴热水的军团菌污染状况进行调查分析,提出相应的对策。方法采集广州市48间亚运接待宾馆酒店淋浴热水108份,按照卫生部《公共场所集中空调通风系统卫生规范》2006年附录A进行军团菌的检测。结果 108份淋浴热水中,10份淋浴热水共检出11株嗜肺军团菌,阳性率为9.3%;其中LP1、LP3、LP6血清型分别占45.5%、18.2%、36.4%;三星级、四星级和五星级宾馆酒店淋浴热水军团菌的阳性率分别为15.2%、11.1%和2.6%。结论广州市亚运接待宾馆酒店的淋浴热水存在嗜肺军团菌的污染,建议定期清洗消毒淋浴设施,并加强对淋浴热水中军团菌的监测,以防范军团菌病的发生。     

深圳市中央空调冷却塔军团菌核糖体基因型研究

军团病(Legionnaires' disease)是由军团菌感染引起的以肺炎为主要症状的急性呼吸道传染病,嗜肺军团菌(Legionella pneumophila,Lp)是引起军团病的主要菌群。目前研究发现军团菌广泛存在于自然界的土壤和各种水环境中,中央空调冷却塔是军团菌主要的传染源,近30年来军团病大的暴发多与中央空调冷却塔被军团菌污染有关。     

淋病奈瑟菌临床菌株PⅠ基因型及PⅠB基因点突变分析

目的分析浙江地区淋病奈瑟菌株外膜孔蛋白PⅠ基因序列及其点突变与耐药性关系，了解本地区淋病奈瑟菌株PⅠA与PⅠB基因分布及其优势基因型。方法采用高保真PCR扩增34株淋病奈瑟菌全长PⅠ基因序列，T-A克隆后测序。根据测序结果，建立多重PCR同时检测113株淋病奈瑟菌PⅠA和PⅠB基因。分析测序菌株PⅠB基因中与耐药性密切相关的G120和A121变异情况，采用二倍琼脂稀释法确定PⅠB菌株的耐药性。结果11株PⅠA^＋淋病奈瑟菌均为ⅠA6血清型。23株PⅠB^＋淋病奈瑟菌中，6株（26．1％）为ⅠB3血清型、5株（21．7％）为ⅠB3／6血清型、2株（8．7％）为ⅠB4血清型、9株（39．1％）为ⅠB3／6-ⅠB2嵌合血清型、1株（4．3％）为ⅠB2-ⅠB4嵌合血清型。所建立的多重PCR可特异和准确地对PⅠA和PⅠB基因分型，检测灵敏度为10雌DNA模板。113株淋病奈瑟菌中，26株（23．0％）和87株（77．0％）分别携带PⅠA和PⅠB基因。经测序的23株PⅠB^＋淋病奈瑟菌中，1株（4．3％）G120和A121均未突变，3株（13．0％）G120或A121突变，2株、8．7％）G120突变但A121缺失，17株（73．9％）双位点突变。22株G120和／或A121突变的PⅠB^＋菌株均对青霉素和四环素耐药。结论所建立的多重PCR可用于淋病奈瑟菌PⅠA和PⅠB基因的分型检测。本地区流行的淋病奈瑟菌主要携带PⅠB基因。ⅠA6为PⅠA菌株优势血清型。PⅠB菌株以ⅠB3／6-ⅠB2嵌合、ⅠB3和ⅠB3／6血清型常见，但主要为耐药性G120和／或A121突变株。     

嗜肺军团菌在武钢青山地区的发现及其检测和防治对策

军团病是因1976年在美国费城退伍军人年会中首次发现急性发热性肺部疾患而被命名的。患者以呼吸道感染为主，死亡率较高。嗜肺军团菌（Legionella pneumophila，Lp）是军团菌属的代表种，是近20年来新发现的传染病之一。我们对武钢青山地区10家单位使用1个月后中央空调系统冷却塔水中的嗜肺军团菌进行检测调查，首次发现嗜肺军团菌的存在，并了解其分布特征和菌型特征，制定出相应的防治策略。     

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     

广州地区肺炎住院患儿感染肺炎链球菌的血清型分布及耐药性研究

目的了解广州地区肺炎住院的患儿感染肺炎链球菌的耐药性及血清型分布情况。方法用吸痰法采集患儿痰标本进行涂片，革兰氏染色镜检，合格痰标本划线接种血平板，用E-test法检测分离到的肺炎链球菌对青霉素、阿莫西林、头孢曲松、头孢呋辛、亚胺培南、氧氟沙星、万古霉素、红霉素和克林霉素9种药物的耐药性，采用K-B法检测四环素、复方新诺明的耐药性，并采用荚膜肿胀技术对分离到的79株肺炎链球菌进行血清分型。结果79株肺炎链球菌中青霉素耐药肺炎链球菌（PRSP）11．4％，青霉素中介肺炎链球菌（PISP）77．2％，青霉素敏感肺炎链球菌（PSSP）11．4％，对红霉素、克林霉素的耐药率分别为100％和93．7％，对阿莫西林、氧氟沙星、万古霉素的耐药率均为0，对头孢曲松、头孢呋辛、亚胺培南的耐药率分别为3．8％、72．2％、2．5％。79株肺炎链球菌中只有1株PSSP仅对红霉素耐药．78株肺炎链球菌对两种以上药物耐药．多重耐药率为98．7％（78／79），同时对克林霉素、红霉素、四环素、复方新诺明耐药的菌株65株，占82．3％（65／79）。79株肺炎链球菌血清型分别为19F（70．9％），23F（16．5％），6B（5．1％），4（2．5％），15B（2．5％），不能分型（2．5％），7价疫苗涵盖率为94．9％（75／79）。结论广州地区肺炎住院患儿肺炎链球菌对大环内脂类抗生素红霉素和林可酰胺类抗生素克林霉素耐药情况严重，对二代头孢菌素头孢呋辛耐药率居高，临床治疗儿童肺炎链球菌感染的肺炎应首选阿莫西林和三代头孢菌素。7价疫苗覆盖率高。预防儿童肺炎链球菌感染所致的肺炎，采用7价疫苗可以达到很好效果。     

两岁以下婴幼儿重型颅脑损伤的临床特点与治疗

目的：探讨两岁以下婴幼儿重型颅脑损伤的临床特点与有效治疗方法。方法：2007年1月至2011年12月期间，我科收治两岁以下婴幼儿重型颅脑损伤患儿24例，其中合并其他部位损伤10例。外伤后均有程度不等的意识障碍，改良GCS评分为3～8分，其中5分以下6例，5～8分18例；呕吐13例，瞳孔变化9例，抽搐或癫痫发作9例。保守治疗16例，手术治疗8例。采用格拉斯哥预后评分（GOS）标准评价治疗效果。结果：24例患儿出院时疗效良好率66．7％（16／24），死亡率16．7％（4／24）；出院后12个月疗效良好率达70．8％（17／24），死亡率达20．8％（5／24）。入院时GCS改良评分与出院后GOS评分之间呈显著正相关。结论：两岁以下婴幼儿有其特有的生理特点，颅脑损伤后意识障碍、生命体征变化、呕吐、癫痫发作等全身症状严重，对失血敏感，治疗有其特殊性，积极治疗可望改善预后。     

Commercial potting soils as an alternative infection source of Legionella pneumophila and other Legionella species in Switzerland

Legionella spp. are pathogens that can cause Legionnaires' disease in humans through inhalation of contaminated aerosols. The principal reservoir for these microorganisms is water, but Legionella spp. have been isolated from composted vegetable and plant material, and from many potting mixes as well. In Australia, there have been several cases of Legionnaires' disease in which Legionella longbeachae has been isolated from potting soils. In Switzerland, the source of infection cannot always be identified as water or cooling towers: therefore, we have investigated 46 commercially available potting soils in Switzerland to determine the presence of Legionella spp. We were able to detect Legionella spp. in 45.7% (21/46) of the potting soil samples analysed by culture. Legionella pneumophila was present in 19.6% (9/46) of the samples and L. pneumophila serogroup  1 in 6.5% (3/46). Quantification by both culture and quantitative real-time PCR revealed high concentrations of legionellae in potting soils, ranging between 10³ CFU/g and 10⁵ CFU/g and 10⁴ genomic units (GU)/g and 10⁶ GU/g, respectively. Thus, potting soils may represent an alternative reservoir for Legionella spp. in Switzerland.     

Isolation of Legionella spp. from cooling tower water and the effect of microbicides

Legionella spp., the causative organism of legionnaires' disease, were isolated from more than 80% of water samples in cooling towers before washing. Therefore, we evaluated the effect of microbicide treatment of cooling tower water on Legionella spp., other bacteria and protozoa. 2-Bromo-2-nitropane-1,3-dial, 2,4-dibromo-5,5-dimethylhydantoin or silver nitrate-treated silica gel was added to cooling tower water. The isolation rate of Legionella spp. in the cooling tower water was 50% after microbiocide treatment with 2-bromo-2-nitropane-1,3-dial being the most effective. The microbicide treatment had no effect on other bacteria or protozoa. These findings indicated the importance of regular washing and water exchange of cooling tower water with microbicide treatment.     

Legionella anisa, a possible indicator of water contamination by Legionella pneumophila

Legionella anisa is one of the most frequent species of Legionella other than Legionella pneumophila in the environment and may be hospital acquired in rare cases. We found that L. anisa may mask water contamination by L. pneumophila, suggesting that there is a risk of L. pneumophila infection in immunocompromised patients if water is found to be contaminated with Legionella species other than L. pneumophila.     

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.     

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.     

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.     

Evaluation of a real-time PCR hybridization assay for rapid detection of Legionella pneumophila in hospital and environmental water samples

A real-time LightCycler assay for Legionella pneumophila was evaluated with 120 water samples potentially contaminated with PCR inhibitors. Results were obtained within five hours, with a detection limit equivalent to 800 cells/L. However, 11 of 22 culture-positive samples containing < 100 CFU/L were also positive by LightCycler assay, indicating the presence of significant numbers of non-viable cells. Following extraction, amplification inhibitors remained in four culture-positive samples, but only one contained > 800 CFU/L. The assay seemed suitable for rapidly screening large sample numbers for heavy contamination with L. pneumophila , but conventional culture should continue to be used to detect low contamination levels.     

Paleoepidemiologic investigation of Legionnaires disease at Wadsworth Veterans Administration Hospital by using three typing methods for comparison of legionellae from clinical and environmental sources.

Multilocus enzyme electrophoresis, monoclonal antibody typing for Legionella pneumophila serogroup 1, and plasmid analysis were used to type 89 L. pneumophila strains isolated from nosocomial cases of Legionnaires disease at the Veterans Administration Wadsworth Medical Center (VAWMC) and from the hospital environment. Twelve L. pneumophila clinical isolates, obtained from patients at non-VAWMC hospitals, were also typed by the same methods to determine typing specificity. Seventy-nine percent of 33 VAWMC L. pneumophila serogroup 1 clinical isolates and 70% of 23 environmental isolates were found in only one of the five monoclonal subgroups. Similar clustering was found for the other two typing methods, with excellent correlation between all methods. Enzyme electrophoretic typing divided the isolates into the greatest number of distinct groups, resulting in the identification of 10 different L. pneumophila types and 5 types not belonging to L. pneumophila, which probably constitute an undescribed Legionella species; 7 clinical and 34 environmental VAWMC isolates and 2 non-VAWMC clinical isolates were found to be members of the new species. Twelve different plasmid patterns were found; 95% of VAWMC clinical isolates contained plasmids. Major VAWMC epidemic-bacterial types were common in the hospital potable-water distribution system and cooling towers. Strains of L. pneumophila which persisted after disinfection of contaminated environmental sites were of a different type from the prechlorination strains. All three typing methods were useful in the epidemiologic analysis of the VAWMC outbreak.