四川省钩端螺旋体病高发县报告病例的准确性调查研究北大核心CSCD

目的了解四川省钩端螺旋体病高发地区报告钩体病例准确性,为进一步控制钩体病流行提供科学依据。方法采集2012年钩体病高发地区报告临床诊断病例血清进行钩体菌培养,血清学诊断,对钩体病检测结果为阴性的标本进行乙脑、出血热和立克次体检测。结果共采集87例报告病例的血清标本,培养出4株钩体菌,钩体菌培养阳性率为4.6%。采集64份病例急性期和恢复期双份血清进行钩体病检测,其中54例阳性,阳性率为84.4%,采集23份病例单份血清,其中6份钩体菌检测阳性,阳性率为26.1%。对27份钩体检测阴性标本进行乙脑ELISA检测,阳性1例,出血热、立克次体检测结果均为阴性。结论四川省钩体病高发地区临床诊断钩体病准确性较高,但依然需要加强病例双份血清采集和检测。     

抗体捕捉酶联免疫吸附法在流行性乙型脑炎早期诊断上的应用

用MacELISA测定了138份临床疑似乙脑病人的血清标本,并与HI试验进行了比较。MacELLSA能特异、敏感地检出乙脑患者发病第2天的IgM抗体,阳性率为55.56％。第4、5病日,IgM抗体阳性率高达92.86％。10例正常人和10份流行性出血热病毒抗体阳性的血清标本无一份阳性。     

山东省沂源县无形体病实验室调查分析

目的通过山东省沂源县人粒细胞无形体及其他立克次体病疫源地小样本试点调查,为全面深入开展立克次体病调查提供数据支持.方法对2004、2005、2006年该地区调查收集的26例可疑患者标本进行血清IgG抗体及PCR回顾性检测分析.2007年7月进一步现场流行病学调查,采集当地48名正常人、10只山羊及8只家狗的血标本以及170只蜱标本进行血清学及分子生物学检测分析.结果 26例发热患者中,8例明确诊断为人粒细胞无形体感染,6例为可疑病例.当地正常人群人粒细胞无形体IgG抗体阳性率为26.7%,人单核细胞埃立克体IgG抗体阳性率为1.8%.检测的10份家养黑山羊血清标本9份阳性.8份家狗标本中,人粒细胞无形体抗体7份阳性,而人单核细胞埃立克体血清抗体全部阳性.来自发热患者、山羊及媒介蜱标本PCR扩增16S rRNA及部分序列分析结果显示高度同源性(100%).结论沂源县农业人群普遍存在人粒细胞无形体、人单核细胞埃立克体及其他立克次体隐性感染状况.当地主要优势蜱种为长角血蜱及嗜群血蜱,两者可能是人粒细胞无形体的重要传播媒介.黑山羊可能是人粒细胞无形体的重要动物宿主,而家狗可能是人粒细胞无形体及人单核细胞埃立克体两者兼顾的重要动物宿主.     

贵州省2010-2014年钩端螺旋体病流行特征分析

目的分析贵州省2010-2014年人间钩端螺旋体(简称钩体)病流行病学特征与鼠类动物带菌情况,为钩体病的防控提供科学依据。方法收集2010-2014年贵州省钩体病例报告数据,利用描述性流行病学方法进行分析。对钩体疑似病例标本进行血清学检测和菌株培养和鉴定。采用夹夜法捕获老鼠对贵州省钩体病高发地区进行鼠类宿主动物及带菌情况进行监测,分析鼠间带菌情况与人间疫情的相关性。结果2010-2014年贵州省报告钩体病人共73例,死亡10例,病死率13.70%。疫情主要集中在黔东南州,5年合计报告病例数52例占全省同期病例的71.23%。6—9月份达到发病高峰。发病人群以农民为主,占总病例数的69.86%。检测钩体疑似病人血清样本18份,其中阳性7份,阳性率为38.89%,均为黄疸出血群抗体。未从钩体疑似病人分离到钩体菌株。鼠间动物监测有效布夹数6 750夹次,共捕鼠646只,鼠密度9.57%,黑线姬鼠密度呈上升趋势,共分离钩体菌株56株,全部分离自黑线姬鼠,带菌率8.67%,经PCR检测均为黄疸出血群钩体。结论贵州省2010—2014年间钩体疫情主要集中在黔东南州,黑线姬鼠为主要带菌鼠类动物,带菌群别为黄疸出血群,与钩体病人血清抗体群别相匹配,提示黑线姬鼠带菌与人间疫情存在相关性。     

江西上高钩端螺旋体病主要传染源（耕牛）和主要流行菌群（七…

本研究分离培养了４４３份病人及疑似病人血清标本，钩端螺旋体阳性率１８．５％，其中２０６份标本ＰＣＲ产物凝胶电泳检测阳性率１４．６％，Ｓｏｕｔｈｅｒｎ杂交阳性率２０．９％．６３头猪肾钩体培养阳性率４．８％，１２３只鼠肾钩体培养阳性率４．０％。２２５份牛尿钩体培养阳性率６．２％，ＰＣＲ产物凝胶电泳阳性经１４．０％，Ｓｏｕｔｈｅｒｎ杂交阳性率１５．８％。     

江西上高钩端螺旋体病主要传染源(耕牛)和主要流行菌群(七日热群)研究

本研究分离培养了443份病人及疑似病人血液标本,钩端螺旋体（钩体）阳性率18.5％。其中206份标本PCR产物凝胶电泳检测阳性率14．6％,Southern杂交阳性革20．9％。63头猪肾钩体培养阳性率4．8％,123只鼠肾钩体培养阳性事4.0％。255份牛尿钩体培养阳性车6．2％,PCR产物凝胶电泳阳性率14．0％,South-ern杂交阳性率15．8％。从病入分离的构体属七日热（七日热和棉兰型）、澳洲、流感伤寒、爪哇、赛罗、拜伦和明尼7个血清群8个血清型,以七日热群为主,占52．7％,其次是澳洲群,占20．0％。从牛尿分离的构体属七日热群七日热型和澳洲群澳洲型,各占50％。来自猪的构体属波摩那和澳洲群,鼠的钩体为爪哇、澳洲及拜伦3个群。结果显示,牛带菌率高,所带钩体血清群型与人间流行钩体主要群型一致。鼠和猪带苗率低,所带菌群与人间流行的主要菌群不相符。以上表明,耕牛是上高县钩体病的主要传染源,主要流行菌型是七日热群（型）。也说明耕牛是我国七日热钩体病主要传染源。这些结果的发现有利于使用对型组合钩体菌苗,控制相应传染源。     

云南省西双版纳州鼠型斑疹伤寒暴发的调查(英文)

目的 阐明云南省西双版纳州鼠型斑疹伤寒的流行状况。方法 收集云南省鼠型斑疹伤寒病例资料。于2011年6-9月在西双版纳地区采集鼠型斑疹伤寒临床诊断病例的急性期和恢复期血清,并在发病地区的居民区捕鼠,采集鼠血液和脾脏标本。用间接免疫荧光试验检测患者和鼠血清中的鼠型斑疹伤寒立克次体的IgM和IgG抗体,用实时荧光PCR试验检测急性期病人血块和鼠脾脏中的鼠型斑疹伤寒立克次体热休克蛋白基因(groEL gene)。结果 2004-2011年,云南省共报告鼠型斑疹伤寒病例8 361例,所有州(市)均有病例报告,其中西双版纳州发病数和发病率最高(86.6/10万),占全省总病例数的73.1%(6 111/8 361)。发病数具有逐年上升之势,全年均有病例发生,6-10月为主要流行期。2011年云南省共报告鼠型斑疹伤寒1 369例,其中西双版纳州报告病例1 157例,发病率为102.10/10万,病例数占云南省病例数的84.51%。勐海县、勐腊县和景洪市分别占全州病例数的79.95%(n=925, 278.74/10万)、18.06%(n=209,74.10/10万)和1.99%(n=23,4.42/10万),勐海县发病率显著高于勐腊县(χ²=346.3, P<0.001)和景洪市(χ²=1369, P<0.001)。对2011年的80例病人进行了实验室检测,在80例急性期病人血清标本中有 63例为IgM抗体阳性;75例病人双份血清标本中有61例的恢复期血清滴度高于急性期4倍及以上;80例患者急性期血块中有8例为PCR阳性。依据实验室诊断标准,74例被确诊为鼠型斑疹伤寒,其中血清学诊断73例(包含7例同时为分子诊断),分子诊断1例,临床诊断和实验室检测符合率为92.50%(74/80)。黄胸鼠血清鼠型斑疹伤寒立克次体IgG抗体阳性率为14.0% (14/100),脾脏的PCR阳性率为9.0% (9/100)。结论 西双版纳州存在较为严重的鼠型斑疹伤寒流行,其中勐海县流行最为严重。     

2021年贵州省一起人间钩端螺旋体病疫情的分子流行病学调查分析

目的 对2021年贵州省一起疑似钩端螺旋体病(钩体病)疫情开展病原学调查和分子流行病学分析,为病例的确诊和疫情防控提供科学依据。方法 对收集的疑似钩体病例全血样本进行分离培养和致病性钩体特异性PCR检测,同时采用显微镜凝集试验(MAT)对病例血清进行钩体抗体检测。采用“夹夜法”捕捉疫区鼠类宿主动物,取肾脏进行钩体分离培养,采用致病性钩体特异性PCR进行检测,进一步采用血清群特异PCR进行鉴定。应用多位点序列分析(MLST)对本起疫情菌株进行基因分型,构建聚类分析图谱,分析其与国内菌株间的遗传和进化关系。结果 3份疑似钩体病例样本经分离培养均为阴性,MAT试验分别为黄疸出血群、秋季群和爪哇群抗体阳性,经致病性钩体特异性PCR检出1例阳性。从鼠类宿主动物样本中分离出6株疑似钩体菌株,经鉴定分别为黄疸出血群(5/6)和秋季群菌株(1/6)。MLST分析显示,鼠类宿主动物携带的5株钩体为ST1型(83.33%),另1株为ST129型(16.67%)。聚类分析显示,鼠类宿主动物携带钩体与四川省、湖南省、江西省和贵州省往年菌株存在同一ST型。结论 本起疫情是由致病性钩体感染引起,优势血清群和ST型...     

ELISA双抗原夹心法检测内脏利什曼病抗体的研究

目的观察rK39抗原酶联免疫吸附试验双抗原夹心法(ELISA夹心法)检测内脏利什曼病抗体用于内脏利什曼病诊断与宿主动物感染调查的可行性。方法采用ELISA夹心法与rK39免疫层析试条法(ICT)同步检测内脏利什曼病抗体。结果 5种家畜血清229份,7种鼠类标本238份,ELISA夹心法和rK39 ICT法抗体检测均阴性;接种利什曼原虫灰仓鼠、草原兔尾鼠标本33份,13份阳性,阳性率39.4%;塔里木兔标本119份,阳性7份,阳性率5.9%;非疫区儿童血清29份,全部阴性;疫区无内脏利什曼病症状人血清250份,4份两种方法均阳性,阳性率均为1.6%;住院内脏利什曼病病人血清67份,两种方法的阳性率分别为68.7%和67.2%。共检测人和其他动物标本965份,两种方法的阳性符合率98.6%,阴性符合率99.9%。ICT相同显色等级的阳性标本,ELISA跨10个滴度。结论 ELISA夹心法可检测多种动物内脏利什曼病抗体,抗体滴度具有定量意义。该方法适用于内脏利什曼病诊断、疗效观察和流行病学调查。     

2009-2012年河北鼠型斑疹伤寒流行概况及实验室调查分析

目的 对2009-2012年河北省斑疹伤寒高发地区临床报告病例进行实验室调查分析。方法 按我国卫生部《流行性和地方性斑疹伤寒诊断标准》(WS 215-2008)在临床可疑病例高发地区设立哨点监测医院并收集病例。临床病例进行个案登记并进行发病急性期及恢复期血液标本的收集。实验室检测按WHO推荐的立克次体血清学诊断方法-间接免疫荧光法检测病人血清IgM和IgG特异抗体。以急性期血液DNA为模板,采用巢氏PCR扩增斑疹伤寒群立克次体热休克蛋白基因groEL基因并测序。通过NCBI Blast平台同源比较序列并使用DNA_STAR软件进行遗传进化分析。结果 本次调查在辛集市、定州市和迁安市共收集877例临床报告病例,2009年报告病例数最多(441例),迁安市病例数最多(510例)。急性期血清莫氏立克次体IgM抗体阳性率为 72.8%(73/101),IgG阳性率为 78.2%(79/101)。12人通过血清IgG抗体4倍升高或降低明确诊断。PCR扩增阳性率为21.7%(22/101),成功测序的17人groEL(209 bp)基因100%同源,与其他地区序列存在较大变异,与标准株R.typhi Wilmington (AY191590)核苷酸97.0%,氨基酸只有93.9%同源,进化分析与其他菌株遗传关系较远。结论 河北省辛集、定州和迁安地区存在鼠型斑疹伤寒流行。进一步调查传播媒介及宿主对该病的预防控制具有重要公共卫生意义。加强该病临床诊断及鉴别诊断十分必要。     

Comparison of a slide agglutination test, LeptoTek Dri-Dot, and IgM-ELISA with microscopic agglutination test for Leptospira antibody detection

A slide agglutination test (SAT), LeptoTek Dri-Dot and IgM-ELISA were compared with a microscopic agglutination test (MAT) for the detection of Leptospira antibodies. Paired sera from 10 patients whose leptospirosis was clinically suspected and diagnosed by MAT, were evaluated in this study. Our data, especially from acute samples, demonstrate the SAT and Dri-Dot were more sensitive as initial screening tests than MAT. IgM-ELISA has an advantage over MAT, SAT, and Dri-Dot since the results can be interpreted from a single serum testing if the results of the test are positive. Eight of the ten cases could be diagnosed by IgM-ELISA. Our data suggest that IgM-ELISA may be used for the diagnosis of leptospirosis. However, the agglutination test is useful for screening and for secondary infection cases for which IgM antibodies may be undetectable. MAT can be performed as a reference test and when information regarding the causative serovar is required.     

Leptospirosis in northern India: a clinical and serological study

A total of 400 serum samples collected from patients, clinically suspected of leptospirosis, were evaluated for antibodies by LEPTO dipstick and microscopic agglutination test (MAT). Twenty of these patients (5%) had serological evidence of leptospirosis. Leptospira interrogans serovars Autumnalis and Icterohaemorrhagiae, Canicola and Javanica were serogroups recorded serologically. Fever and jaundice were the most common clinical presentations. Male preponderance was seen in the leptospirosis cases. Outdoor activities, agricultural activities, contact with animals were significantly associated with seropositivity for Leptospira. This study highlights that leptospirosis is a significant health problem in northern India, though grossly under reported due to the absence of routine laboratory diagnostic facilities for this disease.     

Introduction of a rapid dipstick assay for the detection of Leptospira-specific immunoglobulin m antibodies in the laboratory diagnosis of leptospirosis in a hospital in Makassar, Indonesia

An easy, rapid and robust dipstick assay for detection of leptospira-specific immunoglobulin M (IgM) antibodies was evaluated on 403 patients admitted for hospitalization because of fever. The clinical symptoms and signs of 35 patients were consistent with leptospirosis. The final diagnosis for the remaining patients was as follows: 136 with typhoid fever, 82 with hepatitis, 74 with malaria, 48 with infections of the respiratory tract, and 20 with fever of unknown origin. The clinical diagnosis of leptospirosis was confirmed for 24 (68.6%) patients by the combined results of the microscopic agglutination test (MAT), the reference test for leptospirosis, and of IgM ELISA, a standard laboratory test for the serodiagnosis of leptospirosis. In addition, serum specimens from 8 (2.2%) patients with a final clinical diagnosis other than leptospirosis were found to be positive in MAT and/or IgM ELISA. Compared with the results of MAT and IgM ELISA a sensitivity of 91.6% and specificity of 93.6% was calculated for the dipstick assay. Most of the serum samples from the laboratory confirmed patients gave a moderate to strong staining intensity of the antigen band of the dipstick and were easy to read. The results demonstrate that the dipstick assay is convenient to use and allows the rapid and accurate confirmation of patients with clinical suspicion of leptospirosis in areas where the disease is endemic.     

聚合酶链反应检测致病性钩端螺旋体DNA的研究

以黄疸出血型ＲＧＡ菌株对所有致病性钩端螺旋体特异的ＤＮＡ克隆片段序列为基础合成聚合酶链反应引物Ｇ１Ｇ２。用该对引物扩增各群型致病性钩端螺旋体微量ＤＮＡ，均获阳性结果，对非致病性钩端螺旋体和其他细菌不扩增。纯化的钩端螺旋体ＤＮＡ５ｐｇ经聚合酶链反应后，琼脂糖凝胶电泳可以目测。用地高辛配基标记的特异性探针可检测到５ｆｇ即相当于１条钩端螺旋体ＤＮＡ量的扩增产物。将该对引物扩增早期钩体病患者血清标本，阳性率为８２％，显著高于ＭＡＴ和ｄｏｔ－ＥＬＩＳＡ阳性率。由此表明，ＰＣＲ技术是灵敏、特异、快速的钩端螺旋体病早期诊断方法，还可用于流行病学调查。     

Retrospective serosurvey of leptospirosis among patients with acute febrile illness and hepatitis in Egypt

The epidemiologic status of leptospirosis in Egypt has not been well defined because of difficulties in disease diagnosis. A retrospective study was conducted to detect leptospiral antibodies among undiagnosed acute febrile illness (AFI) and hepatitis cases. Approximately 16% of both AFI (141/886) and acute hepatitis (63/392) cases showed seroreactivity to Leptospira IgM by ELISA and microscopic agglutination test (MAT). Canicola, Djasiman, Grippotyphosa, Pyrogenes, Icterohemorrhagiae, and Pomona were the most commonly reactive serovars among patients with AFI. Djasiman, Grippotyphosa and Icterohemorrhagiae were the most reactive among patients with acute hepatitis. This study represents the first systematic report of Leptospira associated with patients with AFI and hepatitis in Egypt. Physicians need to have increased awareness about the importance of leptospirosis in the differential diagnosis of AFI and acute hepatitis in Egypt. In addition, laboratory capacity should be developed at fever hospitals to diagnose leptospirosis.     

Leptospira borgpetersenii from free-living white-eared opossum (Didelphis albiventris): First isolation in Brazil

Leptospirosis is a zoonotic disease that occurs all over the world, caused by bacteria of the genus Leptospira. Marsupial and didelphidae families are considered susceptible to infection caused by a wide range of Leptospira serovars for which they serve as reservoirs. Thirty-three free-living white-eared opossums (Didelphis albiventris) were captured in Southern Brazil and bodily fluids were collected. From the urine samples it was possible to obtain an isolate identified as Leptospira borgpetersenii by rpoB gene sequencing and belonging to serovar Castellonis by Multilocus Variable-Number Tandem-Repeat Analysis. This is the first report of the isolation of Leptospira spp. from the white-eared opossum in Brazil. In addition, the new strain was also virulent in the hamster model of lethal leptospirosis. The microscopic agglutination test (MAT) was used for detecting the presence of antibodies against Leptospira spp. in white-eared opossum, human, cattle and canine sera using a panel of 59 Leptospira strains that included the new isolate. The inclusion of the new strain in the MAT battery increased the MAT sensitivity for canine sera. These findings suggest that the white-eared opossum is an important reservoir of pathogenic Leptospira spp.     

Use of locally isolated saprophytic Leptospira strain for serological testing of human leptospirosis.

A saprophytic Leptospira isolate recovered from tap water was utilized for serological testing. One hundred-twenty Serum samples comprising 55 cases from PUO/febrile jaundice and 65 samples from apparently healthy individuals were tested by MAT and HA using this environmental saprophytic strain and the results compared with that of Leptospira biflexa semaranga patoc, the standard saprophytic strain commonly employed for sero-diagnosis of leptospirosis. The MAT data showed 96.4 per cent correlation between the two strains. Similarly, the HA results were matching to the extent of 94.5 per cent. Results, therefore, suggest that local saprophytic Leptospira strain may serve as a substitute to serovar patoc for serodiagnosis of leptospirosis.     

Etiology of acute, non-malaria, febrile illnesses in Jayapura, northeastern Papua, Indonesia

We conducted a prospective, inpatient fever study in malaria-endemic Papua, Indonesia to determine non-malaria fever etiologies. Investigations included malaria blood films, blood culture, paired serologic samples analysis for dengue, Japanese encephalitis, leptospirosis, scrub typhus, murine typhus, and spotted fever group rickettsia. During 1997-2000, 226 patients (127 males and 99 females) 1-80 years of age (median age = 25 years) were enrolled. Positive blood cultures (n = 34, 15%) were obtained for Salmonella Typhi (n = 13), Escherichia coli (n = 8), Streptococcus pneumoniae (n = 6), Staphylococcus aureus (n = 5), Streptococcus pyogenes (n = 1), and Klebsiella pneumoniae (n = 1). Twenty (8.8%) patients were positive for leptospirosis by polymerase chain reaction. Eighty (35.4%) of 226 patients had ≥ 1 positive serology, diagnostic for 15 rickettsial and 9 dengue cases. Acid-fast bacilli-positive sputum was obtained from three patients. Most common confirmed (81 of 226, 35.8%)/suspected diagnoses were typhoid fever (n = 41), pneumonia (n = 29), leptospirosis (n = 28), urinary tract infections (n = 20), rickettsioses (n = 19), dengue (n = 17), and meningitis/encephalitis (n = 15). There were 17 deaths, 7 (46.7%) were caused by meningitis/encephalitis. Multiple positive serologic results and few confirmed diagnoses indicate the need for improved diagnostics.     

Leptospirosis in Istanbul, Turkey: a wide spectrum in clinical course and complications

Patients with high fever and multiorgan involvement were investigated for the determination of frequency, clinical course and complications of leptospirosis in Istanbul. Leptospirosis was determined in 22 cases among the 35 hospitalized patients that were pre-diagnosed as leptospirosis according to 'Probable Leptospirosis Diagnosis and Follow-up' form. Among the leptospirosis cases 19 were male and 16 were military staff. Mean age was 35.6 y. Dark field examination (DFE), latex agglutination test (LAG), ELISA IgM, leptospirosis culture (LC) and microscopic agglutination test (MAT) were performed to confirm the diagnoses. The most frequent initial symptoms and findings were fever, fatigue, headache, nausea-vomiting and increased muscle sensitivity. Jaundice was noted only in 2 cases. A 74-y-old female patient died after the recurrence of the disease with severe rhabdomyolysis and pulmonary failure. Sagittal sinus thrombosis, perimyocarditis and chronic renal failure were major complications in another 3 patients. ELISA IgM, LC, DFE, LAG and MAT tests were positive in 68, 72, 82, 100 and 100% of the patients, respectively. As a conclusion, diagnosis of leptospirosis is usually overlooked. Clinical awareness, use of probable leptospirosis diagnosis forms and the application of different laboratory methods in the diagnosis of suspected cases may offer the chance to diagnose the leptospirosis accurately.     

Leptospirosis presenting as acute encephalitis syndrome (AES) in Assam,India

ObjectiveTo establish leptospirosis as a new aetiology of the patients presenting acute encephalitis syndrome (AES).MethodsJapanese encephalitis, West Nile, Dengue and Chikungunya negative samples were tested by IgM capture ELISA for leptospira specific IgM. For further confirmation, the IgM positive samples were subjected to Microscopic agglutination test (MAT). The clinical details and laboratory findings of the positive patients were recorded.ResultsWe report 8 cases of leptospirosis presenting as AES, proven on IgM capture ELISA and confirmed by MAT. Fever (100%) and altered sensorium (62.5%) were two most common symptoms. Low haemoglobin (7.5 ± 2.8) g/dL, elevated blood urea (79.16 ± 46.43) mg/dL, serum creatinine (1.5 ±1.2) mg/dL, SGOT (66.5 ± 14.84) U/L and SGPT (70.5 ± 4.9) U/L were observed in positive patients.ConclusionsThis is the maiden study reporting leptospirosis as a new aetiology of the patients presenting AES. Establishing aetiology is very important for a successful therapy at least in treatable conditions like leptospirosis.