钩端螺旋体病暴发流行特征和病原学、血清学调查分析

目的了解大竹县钩端螺旋体病(简称钩体病)暴发流行特征和病原学、血清学情况,为制定防制措施提供科学依据。方法开展钩体病个案调查,采集病人血液;夹夜法测鼠密度,无菌解剖取双肾;无菌采集猪肾、蛙肾、牛中段尿;将采集的标本进行病原学和血清学分离培养,并进行统计学分析。结果钩体病发病8 356例,发病率为878.71/10万;2 459份病人血液和宿主动物标本中分离培养出钩体菌91株,阳性率为3.70%;病人血液钩体菌检出率明显高于鼠肾、鼠肾高于蛙肾(χ~2=427.48,P0.05),差异有统计学意义;经菌群菌型鉴定为5群8型。七日热群占59.46%,主要从病人血液和蛙肾中分离获得;黄疸出血群为25.68%,主要从鼠肾中分离获得。结论钩体病发病数多,流行强度大,其暴发流行的主要宿主是蛙类和鼠类。     

清远市2004年钩端螺旋体病监测分析

目的 对2004年清远市钩端螺旋体病疫情、人群及宿主动物监测情况进行分析,为制定防治措施提供科学依据.方法 采取疑似病人血、鼠类、水禽类和蛙类动物肾组织分离培养病原体,用MAT法检测健康人、疑似病人、鼠类和水禽类动物血清抗体.结果 报告发生钩体病10例,发病率0.25/10万,死亡1例,病死率10.00%.从鼠肾组织中分离鉴定了5株钩体菌,分离阳性率为5.00%,均为爪哇群.流行前、后期健康人血清和流行期疑似病人血清抗体阳性率分别为13.27%、4.00%和15.79%,菌群以黄疸出血群和七日热群为主,其次为拜伦群和爪哇群.鼠血清抗体阳性率为22.00%,黄毛鼠、板齿鼠、褐家鼠和黄胸鼠的血清抗体阳性率在8.57%～50.00%之间,菌群以秋季热群为主,其次为犬热群.水禽（鸭）血清抗体阳性率为4.00%,菌群为拜伦群和致热群.结论 我市人群钩体隐性感染水平和鼠带菌率均较高,开始出现钩体病发病年龄组后移及钩体带菌优势鼠种由野栖鼠种向家栖鼠种交叉转移的动向,从水禽类动物（鸭）血清中检测出钩体带菌抗体在我市尚属首次.继续加强监测,采取综合性防治措施,可有效地控制钩体病的流行.     

湖南省2005年钩端螺旋体病监测结果与分析

目的 调查了解湖南省钩端螺旋体病的防治监测工作.方法 选择湖南5个县(市)进行传染源带菌情况、自然人群与疑似病人血清学研究,按照国家有关标准进行实验室检测.结果 传染源钩体分离及菌型分布情况对鼠及蛙类标本进行钩体分离培养,分离出15株钩体.其中鼠肾分离培养出阳性标本14株,阳性率3.59%.血清学:检测11份疑似病人血清抗体,确诊5例,确诊率45.45%;525份自然人群血清钩体抗体有12群之多,黄疸出血群里,湘潭县自然人群血清抗体阳性率最高(62.75%),沅江市自然人群血清抗体GMT最高(37.64).结论 从鼠、蛙钩体分离培养中可看出,塞罗群萨克斯可宾型和沅江型是湖南省洞庭湖区主要菌型;黄疸出血群仍是湖南省自然人群主要流行菌型,其次秋季群、澳洲群、波摩那群和流感伤寒群也是湖南省流行菌型;地区重点菌群监测预防工作应加以重视.     

洪灾区钩端螺旋体宿主动物及健康人群感染情况的流行病学研究

目的:了解洪灾区钩端螺旋体宿主动物的带菌情况及人群的免疫水平. 方法:应用柯素夫培养法作牛尿和鼠肾钩体培养.显微凝集试验(microscopic agglutination test,MAT)作钩体及人群钩体感染的血清学分群(型).结果:灾区、临灾区及新建移民点的鼠密度分别为6.95%,6.28%及8.67%;鼠肾钩体培养阳性率分别为4.63%,1.35%和3.13%;牛尿培养阳性率分别为5.88%,5.98%和1.75%;分离的菌株主要为黄疸出血型及犬型;人群钩体抗体阳性率分别为45.91%,62.30%和58.67%,明显高于全省及全国的平均感染率;其主要血清学分型依次为黄疸出血、秋季热、犬型、泼摩那型、巴达维亚型;人群抗体阳性率无年龄差异.结论:3个不同区域的宿主动物鼠和牛中均有钩体带菌,人群钩体隐性感染率水平较高,宿主动物和人群感染的主要血清群相吻合,主要为黄疸出血型.     

我国主要致病钩端螺旋体DNA与OmpL1基因同源性的研究

用＾３２Ｐ标记的ＯｍｐＬ１基因片段与我国１８株钩体进行Ｓｏｕｔｈｅｒｎ杂交分析。此片段与非致病钩体ＰａｔｏｃＩ株、伊利尼细丝体３０５５株无杂交信号；问号钩体中，除爪哇群、塔拉索夫群、曼耗群和明尼群的４株钩 体外，黄疸出血群、犬群、拜伦群、致热群、澳洲群、波摩那群、流感伤寒群、七日热群、巴达维亚群、赛罗群各群钩体参考株ＤＮＡ均有杂交信号。     

应用分子杂交对钩端螺旋体DNA同源性的研究

作者以~(32)P标记的黄疸出血群赖型017株钩体DNA为探针,分别与2个属5个血清群的6株钩体DNA进行分子杂交。结果表明,黄疸出血群赖型017株与同群、型的56601株同源性较高,与秋季群(56606株)、七日热群(56610株)次之,与双曲钩体patoc型Patoc Ⅰ株、细螺旋体属illini细螺旋体同源性很低;~(32)P标记的钩体DNA探针能检测四川地区流行的致病性钩体。     

我国主要致病钩端螺旋体DNA与OmpL1基因同源性的研究

用￣（３２）Ｐ标记的ＯｍｐＬ１基因片段与我国１８株钩体进行Ｓｏｕｔｈｅｒｎ杂交分析，此片段与非致病钩体ＰａｔｏｃＩ株、伊利尼细丝体３０５５株无杂交信号；问号钩体中，除爪哇群、塔拉索夫群、曼耗群和明尼群的４株钩体外，黄疸出血群、犬群、拜伦群、致热群、秋季群、澳洲群、波摩那群、流感伤寒群、七日热群、巴达维亚群、赛罗群各群钩体参考株ＤＮＡ均有杂交信号。     

清远市2000～2003年钩端螺旋体病监测分析

目的：了解清远市2000～2003年钩端螺旋体病人群及宿主动物带菌情况，为制定防治措施提供科学依据。方法：采取病人血、动物脏器分离培养病原体。用MAT法检测健康人、疑似病人和鼠血清抗体。结果：从病人血和动物脏器中分离鉴定了16株钩端螺旋体，分属4种菌群，分别为犬热群、秋季热群、赛罗群和爪哇群。健康人血清和疑似病人血清阳性率分别为29．22％和3．81％，菌群均以黄疸出血群为主。鼠血清阳性率为29．71％，菌群以爪哇群为主，板齿鼠、黄毛鼠、黄胸鼠和褐家鼠的带菌率在26％～50％之间。结论：我市人群钩端螺旋体隐性感染水平和鼠带菌率均较高，从人群中分离到的赛罗群钩端螺旋体在我省尚属首次。继续加强监测，采取综合性防治措施，可有效地控制钩端螺旋体病的流行。     

贵州省2010－2014年间钩体宿主动物带菌监测结果与分析

目的：了解贵州省鼠类宿主动物钩端螺旋体（简称钩体）带菌情况、菌型分布,为钩体病的防控提供技术手段和科学依据。方法：对2010～2014年对贵州省黔东南州钩体疫源地采用夜夹捕鼠法进行鼠间动物监测捕获鼠类,分离钩体后用PCR方法进行鉴定。结果：2010－2014年鼠间动物监测有效布夹数6750夹次,共捕鼠646只（鼠密度9．57％）,以黑线姬鼠最多240只,占总数的37．15％。共分离钩体菌株56株（带菌率8．67％）经PCR检测为黄疸出血群。结论：黑线姬鼠为贵州省钩体病常见的带菌鼠种;黄疸出血群钩体是贵州省鼠间的主要传染源。     

三峡库区兴山县钩端螺旋体宿主动物带菌状况及人群免疫水平分析

目的掌握兴山县钩端螺旋体病(钩体病)宿主动物带菌及人群免疫水平,为进一步做好钩体病的预防与控制提供依据。方法按照《全国钩端螺旋体病监测方案》开展鼠、家畜、青蛙等宿主动物带菌调查及健康人群血清学监测。结果平均鼠密度为1.80%。鼠种构成以黄胸鼠、小家鼠和黑线姬鼠为优势种,平均带菌率为3.75%。共检测病人尿液及家畜、青蛙、稻田疫水等7类样本565份,培养阳性菌株4株,阳性率为0.71%,其中稻田疫水2株,病人尿液、蛙肾各1株,菌株经鉴定黄疸出血群1株,秋季热群3株。人群钩体抗体阳性率平均为28.62%,青壮年阳性率较高,以黄疸出血型、七日热型、流感伤寒型、波摩那型为主。在监测中发现1例在农贸市场感染的经济型钩体病。结论兴山县钩体宿主动物带菌率低,血清学显示人群钩体隐性感染率较低,在山区型钩体自然疫源地存在的情况下,其发病率与社会因素和自然因素密切相关。农贸市场、畜牧养殖区及其周边为经济型钩体病疫源地,病例分析已经得到证实,建议今后应将其纳入预防和监测范围。     

应用低严格度—单引物聚合酶链反应对问号钩体中国参考株的…

Ｇ１、Ｇ２引物是对问号钩体具有特异性引物。分别用Ｇ１或Ｇ２单引物对问号钩体中国参考株进行前４个低严格度循环的ＰＣＲ扩增，扩增带谱显示赖型、犬型、致热型、秋季型、澳洲型、临海型、乌尔夫型、溶血型为一类，而爪哇型、拜伦型、波摩那型、七日热型、巴叶赞型塔拉索夫型、曼耗Ⅱ型是不与以上赖型等血清型本为一类，双曲钩体ａｔｏｃ型及伊利尼细丝体伊利尼型的扩增带谱与问号钩体截然不同。应用苯酚法提取的高纯度钩体ＤＮＡ     

Monoclonal antibodies to Leptospira interrogans serovar pomona

Three monoclonal antibodies produced against Leptospira interrogans serovar pomona have been studied for their diagnostic usefulness. All three monoclonals reacted strongly in the enzyme-linked immunosorbent assay and indirect fluorescent antibody test with serovar pomona and did not react with serovars grippotyphosa, canicola, icterohaemorrhagiae and hardjo.     

用限制性核酸内切酶分析钩端端螺旋体DNA的研究

For the application of restriction endonuclease analysis in typing and identifying leptospira, we selected some serovars and isolates, and analysed preliminarily their DNA with four restriction enzymes, EcoR I, Bgl II, Hha I, and Hind III. The DNA samples were isolated from the reference strains and isolates as follows: Serovar lai 56601, 017 (the virulent strain for PDH model of guinea pig), Serovar autumnalis 56606, Serovar manhao II 67020, and isolates 87112 and 87369. Each 2 micrograms of DNA was digested with 20mu of restriction enzyme at 37 degrees C for 2h and electrophoresed in 0.8% agarose gel. The gels were stained in ethidium bromide and photographed with UV light. In our experiments, apparently different restriction patterns of serovar lai 017 were observed with four restriction enzymes. Serovar lai, serovar autumnalis and serovar manhao II showed different patterns with EcoR I, especially in high molecular regions. We also observed in serovar lai 017 a distinct 10.5kb band which was obscure in 56601, the reference strain of serovar lai, after EcoR I digestion. The three serovars showed some delicate differences in Hind III restriction pattern. The two isolates from Apodemus agrarius in Sichuan (1987) 87112 and 87369 had patterns identical to those of serovar lai 56601, 017 with EcoR I, and 87112 also had a pattern identical to 56601, 017 with Hind III. Our results indicate that selected three serovars can be identified by analysis of their DNA with EcoR I and Hind III. It is suggested that restriction endonuclease analysis be a good method in typing and identify leptospira and in studying the differences of special DNA molecules.     

应用分子杂交对钩端螺旋体DNA同源性的研究

Homology of leptospires from different genus, different serogroups were studied with molecular hybridization. Leptospiral DNAs were extracted and purified with phenolchloroform-isoamylalcohol method. Alpha 32P-dCTP was used to label DNA from L. interrogans serogroup icterohaemorrhagiae serovar lai strain 017 as a DNA probe, and hybridized with DNAs of 2 genus, 5 serogroups of leptospires represented by 6 strains on NC filter. Four serogroups of pathogenic leptospires which caused endemic disease in Sichuan Province were also detected by the probe. The results showed that L. interrogans serogroup icterohaemorrhagiae, serogroup autumnalis and serogroup hebdomadis had a high degree of homology while there was a low degree of homology between L. interrogans and L. biflexa and Leptonema illini. Four major serogroups of pathogenic leptospires in Sichuan Province, with their high degree of homology, could be detected by a radiolabelled probe from serogroup icterohaemorrhagiae. Hybridization may be used as a tool for diagnosis of leptospirosis in human beings and animals.     

钩端螺旋体重组DNA探针对致病性黄疸出血群钩体DNA的杂交识别

Two recombinant DNA probes-PLIpso1 (15kb) and PLIEc34 (4kb), derived from Leptospira serovaricterohaemorrhagiae genomic libraries, were applied for the hybridization and identification of 13 strains of Leptospira in serogroup icterohaemorrhagiae. Difference in hybridization signal in combination with the banding pattern provide a good way for identification of serovars and strains. The recombinant DNA, specific to L. serogroup icterohaemorrhagiae, hybridized with a limited number of DNA fragments which had been digested by several restriction endonucleases. The less complex banding pattern and higher sensitivity facilitate characterization of various serovars and strains in serogroup icterohaemorrhagiae. In general the DNA patterns recognized by both probes have extensive genomic homology in same serovar (but still can distinguish strains in same serovar by some unique bands) and apparent difference in various serovars (especially serovars naam, nanxi and honghe). The results indicated that Southern blotting with recombinant DNA probe might provide tools for identification, characterization and analysis of leptospira.     

针对问号钩体赖型外膜的抗原分析

作者采用SDS-聚丙烯酰胺凝胶电泳和免疫印迹技术,对黄疸出血群赖型017株和601株、七日热群七日热型156株、澳洲群澳洲型620株以及塞马伦群帕托克型Patoc Ⅰ株等5株钩体外膜进行分析。检测到具有免疫保护作用和抑制钩体粘附作用的单克隆抗体E_4B_7D_5能特异地识别赖型017株和601株钩体外膜的34.5kd和39.5kd两条蛋白带,而同其余3株钩体外膜蛋白无反应。由此提示该两条蛋白带可能为钩体外膜保护性抗原。     

Observation of monoclonal antibody E4B7D5 inhibitory effect on leptospiral adherence using scanning electron microscope]

BALB/c mice were immunized intraperitoneally with outer envelopes of serogroup icterohaemorrhagiae lai serovar strain 017 leptospires. Monoclonal antibody (McAb) E4B7D5 against outer envelopes (IgG1, agglutinating titre 1:25,600) was produced by hybridoma technique. Passive immunoprotection experiments have demonstrated the immunoprotection of McAb E4B7D5 against strain 017 leptospires. Effect of McAb E4B7D5 on leptospiral adherence to the surface of normal human pulmonary embryonic fibroblasts was observed by using scanning electron microscope. The results indicated that the leptospiral adherence noted in various agglutinating titre McAb E4B7D5 groups was less frequent than that in the three control groups. It was concluded that the inhibitory effect of McAb E4B7D5 on leptospiral adherence may play a role in the immunoprotection.     

Reemergence of chloramphenicol sensitivity in enteric fever

Background: Enteric fever is a global health problem and rapidly developing resistance to various drugs makes the situation more alarming. Drug sensitivity in Salmonella enterica serovar typhi and Salmonella enterica serovar paratyphi A isolated from 45 blood culture positive cases of enteric fever was tested to determine in-vitro susceptibility pattern of prevalent strains in northern India. Methods: Strains isolated from 45 blood culture positive cases of typhoid and paratyphoid fever over a period of three years were studied and their sensitivity patterns to chloramphenicol, ampicillin, ciprofloxacin, ceftriaxone, nalidixic acid, amikacin and ofloxacin were analysed.     

Leptospirosis in Barbados. A clinical study

A 39-month clinical study of leptospirosis was undertaken at the Queen Elizabeth Hospital, Barbados. Eighty-eight patients had a confirmed diagnosis of the disease during the period. The major serogroups identified were autumnalis (including a new serovar bim), icterohaemorrhagiae, ballum and canicola. The majority of patients presented with jaundice (95%,) anorexia and headaches (85%), fever (76%) and conjunctival suffusion (54%). While abnormal creatinine levels were seen in 49% of patients on admission, only 16% were judged to have had renal failure. The urine to plasma urea ratio showed high sensitivity and specificity in the diagnosis of pre-renal azotemia. Cardiac arrhythmias and myocarditis occurred in 18% of patients and pericarditis in 6%. An elevated serum amylase was found in 65% of cases. The bilirubin level took 5.5 weeks to return to normal. Thrombocytopenia was shown not to be due to a disseminated intravascular coagulation, and a randomised trial of high dose penicillin did not reveal any benefit to jaundiced patients. The overall mortality during the study was 5.7%.     

钩端螺旋体重组DNA探针对致病性黄疸出血群钩体DNA的杂交识别

作者从建立的黄疸出血群钩端螺旋体(下称钩体)DNA基因库中筛选出重组DNA克隆PLIps01(15kb)和PLIEc34(4kb)制备成~(32)P放射性探针。以该二探针对致病性黄疸出血群所属血清型的13个菌株钩体DNA进行Southern印迹分析,结果表明,该二DNA探什对黄疸出血群各株钩体DNA具有强烈的特异性结合,能识别出简洁的DNA带型,并能区分不同血清型以至菌株的DNA带型差别。作者认为,钩体DNA重组探什结合Southern印迹分析是一种灵敏而特异的检测分析方法,可作为诊断、鉴定和分析钩体的工具。