非O1群霍乱弧菌噬菌体-生物学分型

目的：了解用霍乱弧菌噬菌体和溶血性等4项生物学试验对非O1群霍乱弧菌进行噬菌体-生物学分型实际应用意义。方法：用VP1-VP55组弧菌噬菌体对非O1群霍乱弧菌进行噬菌体分型；用溶源性噬菌体敏感试验、溶源性测定、山梨醇发酵和溶血试验对非O1群霍乱弧菌进行生物学分型。结果：236株非O1群霍乱弧菌中能被5组O1群霍乱弧菌噬菌体分型的62株，总分型率为26．3％，其中腹泻病人52株，从业人员10株，分型率分别为22．0％和9．3％，分属于11个噬菌体型，腹泻病人以21、26噬菌体型居多，从业人员无优势型别，对5组O1群霍乱弧菌体均不敏感的174株；62株非O1群霍乱弧菌分属于e、f、k、l等4个生物型，23个噬菌体-生物型在8-31型之间，均属非流行株范畴；对第Ⅳ组噬菌体（10^9／ml）及其常规稀释噬菌体和溶原性噬菌体、溶源性表现为不裂解。结论：非O1群霍乱弧菌进行噬菌体分型，对预测非O1群霍乱弧菌流行具有一定的实用和指导性意义。现成的噬菌体分型方法对非O1群霍乱弧菌进行噬菌体分型存在一定的缺陷，应寻找更适宜于非O1群霍乱弧菌分型的噬菌体用于实际应用。     

温州市1999年霍乱病原学鉴定报告

1999年本实验室收到各县 (市 )站上送霍乱弧菌菌株共133株 ,其中Ｏ13 9群霍乱弧菌 12 2株 ,小川型霍乱弧菌 11株。经血清学、噬菌体弃去生物分型及药敏试验 ,结果发现 :今年的噬菌体分型与药敏试验有新的动向。现报告如下。材料与方法　 (1)菌株来源 :131株来自病人与带菌者 ;1株来自砧板 ,1株来自马桶采样。 (2 )血清及噬菌体 :来自浙江省卫生防疫站防疫科 ,方法按文献〔1〕。 (3)药物敏感试验 :采用琼脂扩散法 ,使用浙江省卫生防疫站生物试剂有限公司的药敏纸片及判断标准。结果　 (1)血清学分型 :133株霍乱弧菌 ,其中Ｏ139群 12 2株 ,占 …     

下城区健康人群粪便中检出O1和O139群霍乱弧菌毒力株和无毒力株

目的：对下城区饮食、服务行业人员粪便标本进行霍乱弧菌监测。方法：2007年1月-2007年9月采集25052名饮食、服务行业从业人员健康体检者粪便标本,分别接种于碱性蛋白胨水增菌后分离4号琼脂。通过培养特性、菌落形态、革兰染色、动力和生化试验等检查,对所分离的疑似霍乱弧菌菌株进行初步鉴定,采用霍乱弧菌O1群及O139群单克隆抗体的玻片凝集试验、噬菌体生物分型、霍乱弧菌ctxA和tcpA基因PCR检测进一步鉴定各霍乱菌株。结果：上述标本中检出O1血清群和O139血清群霍乱弧菌各1株。1株O1血清群霍乱弧菌噬菌体生物分型为1 L,属埃尔托生物型非流行菌株,霍乱弧菌ctxA和tcpA毒力基因PCR检测结果为ctxA阴性,tcpA阳性,判定为有毒力的非流行菌株。另1株O139血清群霍乱弧菌ctxA和tcpA基因PCR检测结果均为阴性,为无毒力非流行菌株。结论：尽管所检出的2株霍乱弧菌均为非流行菌株,但因携带者从事职业的特殊性,仍应引起高度重视。     

呼和浩特市首次霍乱爆发的病原菌鉴定

采集土左旗北什轴乡西红岱村病人和密切接触者粪便９７７ 份样品进行霍乱弧菌检验。对检出的１０ 株可疑菌株进行形态特征、培养特征、生化反应、血清学凝集反应、噬菌体分型等系统鉴定。鉴定结果确认引起呼市首次霍乱爆发的病原菌为霍乱弧菌埃尔托生物型、小川血清型、１ｂ 噬菌体生物型流行株     

海口地区240株非O1群霍乱弧菌血清学分型报告

海口地区２４０株非Ｏ１群霍乱弧菌血清学分型报告海口卫生检疫局（海口５７０２０６）黄廷学黄春德中国药品生物制品检定所陈天寿非Ｏ１群霍乱弧菌在自然界中分布广泛，在病人粪便及外环境中常能检出。大量调查资料表明，该菌不仅能引起人类急性腹泻，而且还可引起胃肠道...     

2005年福建省霍乱弧菌流行菌型特征分析

目的:了解福建省霍乱弧菌流行菌型特征,为控制传染来源提供科学依据。方法:采用血清学分型法、噬菌体-生物分型法及PFGE(脉冲场凝胶电泳)分型法对2005年霍乱疫情中不同来源的142株霍乱弧菌进行分型分析。结果:142株霍乱弧菌血清学、噬菌体-生物分型结果:流行株124株中O1群埃尔托稻叶1d型占95.16%(118/124株);其余菌株为6型以上非流行株。70株霍乱弧菌PFGE分型结果呈现3大类8种带型。结论:2005年霍乱疫情期间患者菌株与福州市闽江水系分离株菌型一致,表明具有高度同源性,而与市场水产品不相关。流行菌型更换而出现的埃尔托稻叶1d型,提示福建省今后仍有发生霍乱流行的可能。     

珠江河口水体霍乱弧菌污染状况调查研究

目的:了解珠江河口水体01/0139群霍乱弧菌的污染情况,提出改善水体霍乱弧菌监测的建议,为采取针对性措施预防控制霍乱的发生和流行提供依据.方法:根据历年霍乱监测资料和疫点分布状况,在珠江水系广州段选择设立24个采样点,每月采集水体标本进行01/0139群霍乱弧菌分离培养,对分离得到的阳性菌株进行血清分型、噬菌体一生物分型、PCR检测毒力基因、K-B法药敏试验.结果:2006年3月～2007年2月共采取862份水体样本,其中有67份检出01或0139群霍乱弧菌,检出率为7.77%,血清型以01群为主,尤以稻叶型占优;01群菌株均为非流行株,2株0139群菌株检出ctx毒力基因;菌株耐药分析发现不同血清型菌株耐药谱有所差异.结论:01/0139群霍乱弧菌在珠江河口水体环境中广泛存在,常年均可从水体中分离获得.必须加强环境水体及海水产品霍乱弧菌监测,及时了解其污染状况,以便采取针对性措施预防控制霍乱的发生和流行.     

嘉兴市霍乱弧菌分离株检测分析

目的比较分析嘉兴市霍乱弧菌菌株的血清型、毒力基因和分子分型特征。方法采用血清学和分子生物学方法对近年来分离到13株霍乱弧菌菌株进行血清型分布、毒力基因(ctxA、ace、zot、tcpA、cri和rtxA)携带和ERIC-PCR分型研究。结果 13株霍乱弧菌菌株,2株为O139群霍乱弧菌,11株为O1群霍乱弧菌,其中小川型9株,稻叶型2株。2株O139群霍乱弧菌菌株携带3种毒力基因;11株O1群霍乱弧菌菌株,除1株携带4种毒力基因外,其余10株携带全部6种毒力基因。ERIC-PCR分型分析显示,不同的霍乱弧菌菌株之间ERIC-PCR型别表现出一定的遗传多样性。结论嘉兴市霍乱弧菌菌株大多携带多种毒力基因,而且来源多样,防控工作任重而道远。     

非01群霍乱弧菌血清分型研究

为研究非 O1群霍乱弧菌的血清型别及分布规律,1987年河北省在3市(秦皇岛、唐山及保定)4地区(廊坊、衡水、沧州及邯郸)进行了非 O1群霍乱弧菌的定点检测,共分离到426株菌。用中国药品生物制品检定所建立的 VBO系统进行血清分型,可分型的有220株,占51.64%,属于52个血清型。其中人源株分型率达85%(51/60),水源株为40.68%(72/177)、泥株为68.75(11/16),水生动物源株为55.79%(77/138)、水生植物源株为60%(9/15)。     

河南省非0—1群霍乱弧菌血清分型的研究

我们采用中国药品生物制品检定所最新研制的霍乱弧菌“O”分型血清81型,首次对河南省非0—1群霍乱弧菌196株进行了血清分型的研究,不同来源的非0～1群霍乱弧菌,除24株未定型者外,可分为37个血清型,分型率为87.8%。以 O_2和O_7型较多。作者认为,O_2血清型为河南省已发现的主要菌型,而且是我省腹泻病患者主要致病菌型。     

广州海珠地区非O1/非O139群霍乱弧菌流行状况调查及生物学特征研究

目的 了解广州海珠地区2001-2009年非O1/非O139群霍乱弧菌的流行、分布状况及生物学特征,为防控该类病原菌引起的急性腹泻病提供科学依据.方法 参照<霍乱防治手册>(第5版)中的检测方法,对本区珠江河水、饮用水、养殖水、各种海(水)产品、市售熟食等外环境标本764份、健康人群标本189份及肠道门诊腹泻患者标本3398份,共计4351份标本进行非O1/非O139群霍乱弧菌的检定.结果 4351份标本共检出非O1/非O139群霍乱弧菌101株,总检出率为2.32%;其中有66株可分型,分属于26个血清型,分型率为65.3%;VBO9、VBO38和VBO76为优势菌.外环境与腹泻患者标本中分离到9株同型别非O1/非O139群霍乱弧菌,提示两者可能存在关联.结论 非O1/非O139群霍乱弧菌在本地区的血清型分布呈现多样性,在人群中有一定的感染率;外环境中该类菌群广泛存在,对人群是一个潜在的危害因素.     

Association of a disease approximating cholera caused by Vibrio cholerae of serogroups other than O1 and O139.

One hundred and six patients suffering from severe dehydrating diarrhoea were studied of whom 36 patients were positive for Vibrio cholerae. Out of 36, 15 were positive for V. cholerae O1, 10 for V. cholerae O139 and 11 for V. cholerae non-O1 non-O139. O1 and O139 were positive for the 301-bp ctxA amplicon and 471-bp tcpA amplicon indicating that the strains possessed toxigenic capability whereas no non-O1 non-O139 strain possessed ctxA or tcpA genes. Post-admission severity of purging and amount of ORS required were less in the V. cholerae non-O1 non-O139 group (P < 0.05) compared to the V. cholerae O1 and O139 groups. It appears from this study that a cholera-like clinical condition can be caused in the absence of CT as exemplified by strains of non-O1 non-O139.     

A high proportion of Vibrio cholerae strains isolated from children with diarrhoea in Bangkok, Thailand are multiple antibiotic resistant and belong to heterogenous non-O1, non-O139 O-serotypes.

Results of a surveillance on cholera conducted with patients seen at the Children Hospital in Bangkok, Thailand from August 1993 to July 1995 are presented. Annually, isolation rates for Vibrio cholerae varied between 1.7 and 4.4% of patients with diarrhoea. V. cholerae O1 serotype Ogawa accounted for between 31 and 47% of patients cultured positive for V. cholerae, whereas the O139 serotype dominated in early 1994 after which it disappeared. Non-O1, non-0139 strains were isolated at similar rates as serotype O1 in 1993 and 1994, but accounted for 69% of V. cholerae culture positive specimens in 1995. However, the annual proportion of the isolation of non-O1, non-O139 strains showed little variation and remained low between 1.0 and 1.3%. Serotyping of 69 epidemiological unrelated non-O1, non-O139 strains produced 37 different O-serotypes. BglI ribotyping of serotypes containing more than two strains demonstrated a high degree of heterogeneity within and between serotypes, except seven serotype O37 strains which showed an identical ribotype suggesting clonality. None of the 69 strains hybridized with a cholera toxin probe and only two strains hybridized with a heat-stable enterotoxin probe. Susceptibility testing to 12 antibiotics showed that 40 of 69 (58%) non-O1, non-O139 strains were resistant to colistin, streptomycin and sulphisoxazole and 28 of 69 (41%) were multiple antibiotic resistant (MAR; > or = 4 antibiotics). Although 26 of 69 (38%) strains contained one or more plasmids, the plasmids were of low molecular weights and did not seem to encode antibiotic resistance. The results of the present study showed that a high proportion of heterogenous MAR V. cholerae non-O1, non-O139 strains were isolated from children at the hospital. With reference to the emergence of V. cholerae O139 in 1992, we suggest that non-O1, non-O139 strains should be monitored carefully to detect new serotypes with a possible epidemic potential, but also to determine the development and mechanism of antibiotic resistance.     

Ribotypes of clinical Vibrio cholerae non-O1 non-O139 strains in relation to O-serotypes

The emergence of Vibrio cholerae O139 in 1992 and reports of an increasing number of other non-O1 serogroups being associated with diarrhoea, stimulated us to characterize V. cholerae non-O1 non-O139 strains received at the National Institute of Infectious Diseases, Japan for serotyping. Ribotyping with the restriction enzyme BglI of 103 epidemiological unrelated mainly clinical strains representing 10 O-serotypes yielded 67 different typing patterns. Ribotype similarity within each serotype was compared by using the Dice coefficient (Sd) and different levels of homogeneity were observed (serotypes O5, O41 and O17, Sd between 82 and 90%: serotypes O13 and O141 Sd of 72; and O2, O6, O7, O11, O24 Sd of 62-66%). By cluster analysis, the strains were divided into several clusters of low similarity suggesting a high level of genetic diversity. A low degree of similarity between serotypes and ribotypes was found as strains within a specific serotypes often did not cluster but clustered with strains from other serotypes. However, epidemiological unrelated O5 strains showed identical or closely related ribotypes suggesting that these strains have undergone few genetic changes and may correspond to a clonal line. Surprisingly, 10 of 16 O141 strains studied contained a cholera toxin (CT) gene, including 7 strains recovered from stool and water samples in the United States. This is to our knowledge the first report of CT-positive clinical O141 strains. The closely related ribotypes shown by eight CT-positive strains is disturbing and suggest that these strains may be of a clonal origin and have the potential to cause cholera-like disease. Despite the low degree of correlation found between ribotypes and serotypes, both methods appears to be valuable techniques in studying the epidemiology of emerging serotypes of V. cholerae.     

A zymovar analysis of Vibrio cholerae isolated in Australia

Zymovar analysis was used to study 50 strains of Vibrio cholerae O1 and 40 strains of V. cholerae non-O1 isolated in Australia. The strains were assigned to 42 zymovars; the O1 strains to 9 types and the non-O1 strains to 33 types, with no overlapping between serovars. All the human O1 isolates, regardless of their ability to produce cholera toxin (CT), and all the CT-producing O1 environmental isolates, were type Z14. The remaining O1 strains and the non-O1 strains belonged to a variety of zymovars, and more than one zymovar was present in some rivers.     

Molecular characterization of Vibrio cholerae O1 and non-O1 from human and environmental sources in Malaysia

A total of 31 strains of Vibrio cholerae O1 (10 from outbreak cases and 7 from surface water) and non-O1 (4 from clinical and 10 from surface water sources) isolated between 1993 and 1997 were examined with respect to presence of cholera enterotoxin (CT) gene by PCR-based assays, resistance to antibiotics, plasmid profiles and random amplified polymorphic DNA (RAPD) analysis. All were resistant to 9 or more of the 17 antibiotics tested. Identical antibiotic resistance patterns of the isolates may indicate that they share a common mode of developing antibiotic resistance. Furthermore, the multiple antibiotic resistance indexing showed that all strains tested originated from high risk contamination. Plasmid profile analysis by agarose gel electrophoresis showed the presence of small plasmids in 12 (7 non-O1 and 5 O1 serotypes) with sizes ranging 1.3-4.6 MDa. The CT gene was detected in all clinical isolates but was present in only 14 (6 O1 serotype and 8 non-O1 serotype) isolates from environmental waters. The genetic relatedness of the clinical and environmental Vibrio cholerae O1 and non-O1 strains was investigated by RAPD fingerprinting with four primers. The four primers generated polymorphisms in all 31 strains of Vibrio cholerae tested, producing bands ranging from < 250 to 4500 bp. The RAPD profiles revealed a wide variability and no correlation with the source of isolation. This study provides evidence that Vibrio cholerae O1 and non-O1 have significant public health implications.     

可疑霍乱弧菌的实验鉴定

目的:对4株可疑霍乱弧菌进行种属鉴定。方法:对可疑菌落在传统血清学和系统生化鉴定的基础上进行细菌的分子鉴定:应用16S rDNA的序列分析进行细菌种属鉴定和实时荧光PCR检测O1、O139群霍乱弧菌。结果:16 s rDNA基因序列分析显示041和067为麦氏弧菌,059和074为霍乱弧菌;荧光定量PCR检测结果说明4株菌都不是O1和O139霍乱弧菌。结论:结合血清凝集和系统生化实验,可以确定041和067为麦氏弧菌,059和074为非O1/非O139霍乱弧菌。对于可疑和难鉴别的霍乱弧菌,不能单纯依靠血清学和表型鉴定方法,需要分子诊断方法,尤其是16SrDNA的序列分析,可以从分子水平为弧菌科细菌的种属鉴定提供更为直接的证据。     

Expanding multiple antibiotic resistance among clinical strains of Vibrio cholerae isolated from 1992-7 in Calcutta, India

Antimicrobial susceptibilities of Vibrio cholerae strains isolated from cholera patients admitted to the Infectious Diseases Hospital, Calcutta, India for 6 years were analysed to determine the changing trends; 840 V. cholerae strains isolated in 1992-1997 were included in this study. Among V. cholerae serogoup O1 and O139, ampicillin resistance increased from 1992 (35 and 70%, respectively) to 1997 (both serogroups 100%). Resistance to furazolidone and streptomycin was constantly high among V. cholerae O1 strains with gradual increase in resistance to other drugs such as ciprofloxacin, co-trimoxazole, neomycin and nalidixic acid. V. cholerae O139 strains exhibited susceptibilities to furazolidone and streptomycin comparable with those of O1 strains. However, after initial increase in resistance to chloramphenicol and co-trimoxazole, all the V. cholerae O139 strains became susceptible to these two drugs from 1995 onwards. Both V. cholerae O1 and O139 remained largely susceptible to gentamicin and tetracycline. V. cholerae non-O1, non-O139 strains, in contrast, exhibited high levels of resistance to virtually every class of antimicrobial agents tested in this study especially from 1995. Kruskal-Wallis one-way analysis showed that V. cholerae O1 Ogawa serogroup exhibited significant yearly increase in resistance to nine antibiotics followed by non-O1 non-O139 and O139 strains to six antibiotics and two antibiotics respectively. Interesting observation encountered in this study was the dissipation of some of the resistant patterns commonly found among V. cholerae non-O1 non-O139 or O1 serogroups to the O139 serogroup and vice versa during the succeeding years.     

The prevalence of Vibrio spp. in drinking water and environmental samples in Vellore South India.

The prevalence of Vibrio cholerae in drinking water, lakes and sewage outfalls during July and August 1996 in Vellore, India was determined. Drinking water samples were collected on single occasions from 12 sites in different geographic areas of the town where cholera had been reported. Samples of water, plankton and sediment were collected from fixed sites at three lakes on three occasions separated by at least 3 days during the course of the study. Samples from open sewers were taken from two representative sites in four areas of the town. Bacteria isolated from samples were identified by standard biochemical tests and isolated strains of V. cholerae tested for their ability to agglutinate O1 and O139 antisera. Water samples from lakes were also tested for the presence of V. cholerae O1 and O139 by fluorescent antibody staining. Non-O1, non-O139 strains of V. cholerae were detected in 41% of drinking water samples and 100% of water, sediment and plankton samples from the test lakes. Eighty-seven per cent of open sewers sampled contained viable non-O1, non-O139 V. cholerae. Fluorescent antibody staining gave positive results for V. cholerae O1 and O139 for all water samples from the three lake sites. Strains of Aeromonas spp. were isolated from 58% of drinking water samples and from 66% of sediment, 77% of plankton and 55% of water samples from lakes. All open sewers sampled contained Aeromonas spp. PCR amplification employing specific primers demonstrated that none of the non-agglutinating V. cholerae isolates contained the ctx operon. The non-O1, non-O139 V. cholerae isolates showed different patterns of antibiotic resistance to ampicillin, ciprofloxacin, chloramphenicol, tetracycline and trimethoprim.     

一例霍乱带菌者携带两株霍乱弧菌的生物学特性研究

目的:对同一霍乱带菌者中分离的2种血清型霍乱弧菌开展生物学特征研究。方法:应用噬菌体生物分型方法、药敏纸片法、PCR方法和脉冲场凝胶电泳试验对两株菌开展研究。结果:两株菌的血清型、噬菌体生物型均不同,耐药性、毒力基因结果相同,脉冲场凝胶电泳分型结果显示带型高度相似。结论:两株菌虽然存在一定差异,但差异非常小,应为同源菌株。