用Western印迹法鉴别布氏菌和O:9型耶氏菌抗体

本文用Western印迹法证实O:9型耶氏菌外膜蛋白中分子量为36、22、12kda等多肽特异于布氏菌,几乎和所有静脉接种、皮下感染O:9型耶氏菌家兔血清抗体反应。用YOMPs为抗原的Western印迹法,结合布氏菌试管凝集试验(STA),可以应用于布氏菌和O:9型耶氏菌血清学鉴别诊断。该技术具有敏感、可靠的特点。     

4头猪耶氏菌感染误诊为布氏菌感染

布病与0∶9型耶氏菌的血清学交叉反应,国外早有报告,国内亦有数篇报告。最近我们发现4份猪血清,按布病常规检测方法判定为布病阳性,经过鉴别试验,确认为耶氏菌感染,类似例子不少,特此介绍,以引起重视。 材料和方法 一、标本来源 在莆田调查布病时,使用布病试管凝集试验法检查猪血清,有4份血清抗体滴度达到1∶100～1∶400,按规定应判为布病阳性。 二、检测方法 1.试管凝集抗原为兰州生物制品研究所产品,在有效期内使用。 2.快速酶斑点试验:在前文的基础上,改用0∶3型耶氏菌外膜蛋白为抗原,布氏菌抗原为全菌可溶性抗原。将这两种抗原滴于同一条膜上,自然凉干后封闭。详细方法和判定见之文献。     

鉴别布氏菌与0：9型小肠结肠炎耶氏菌两种抗体的研究

作者应用经异种抗体吸收的羊种布氏菌１６Ｍ及０：３型小肠结肠炎耶氏菌菌体抗原做ＥＬＩＳＡ对３个种６株布氏菌及Ｙｅ０：９感染的兔血清检测的结果表明，对同种抗原的抗体显示高滴度而对异种抗原的抗体则为阴性反应。这明显优于用１６Ｍ及Ｙｅ０：３外膜蛋白抗原做ＥＬＩＳＡ的鉴别结果。     

从病人标本检出的小肠结肠炎耶氏菌致病力测定

对河南省近年从病人标本中检出的小肠结肠炎耶氏菌进行了致病力测定。体外测毒试验表明,76.9％的O:3和O:9血清型菌株为阳性;体内测毒试验表明,64.7％的O:3、O:9和O:6,30血清型菌株为阳性。实验证明:耶氏菌0:3、0:9和0:6,30为主要致病血清型,个白鼠眼球后注射致死性试验,为耶氏菌体内测毒较好的方法。     

布鲁氏菌与小肠结肠炎耶氏菌血清学鉴别诊断

1969年,芬兰学者Ahvonen等首次报道了布鲁氏菌(简称布氏菌)与0:9型小肠结肠炎耶尔森氏菌(简称耶氏菌)之间存在严重的血清学交叉反应。从而引起了国内外学者极大关注。现已表明,布氏菌与几十种微生物有血清交叉反应,其中以耶氏菌最为严重。目前,我国开展人畜间布氏菌病(简称布病)调查,尤其在布病非流行地区仍采用常规血清学诊断方法,而这些常规方法无法排除布氏菌与其它微生物之间的血清交叉反应。本文自1991年以来,收集我省部分历史布病流行地区家猪、牛、羊和可疑布氏菌感染人群血清,进行布氏菌与耶氏菌血清学鉴别诊断研究,现将结果报道如下。     

布氏菌和0∶9型小肠结肠炎耶氏菌两种抗体鉴别的研究

本文报告一种布病检测的简便方法,以硝酸纤维素膜为固相栽体的免疫斑点试验,使用两种抗原(牛种布氏菌和O:3型耶氏菌)进行测定,其敏感性和特异性均高于常规的布病试管凝集试验,而且方法简便,快速,适于基层推广使用,可以代替试管凝集试验,作为布病监测的首选方法。     

牛种布氏菌与0∶9型小肠结肠炎耶氏菌的鉴别

牛种布氏菌与0∶9型小肠结肠炎耶氏菌(以下简称耶氏菌)的交叉反应,已为许多学者所证实,曾经使用过各种方法包括凝集试验、补体结合试验、抗球蛋白试验、间接血凝试验、2—ME 试验、免疫荧光技术、ELISA 和放射免疫等,企图加以鉴别,均未获得满意的简单方法。本文在研究这两种菌抗原差异性的基础上,提出一个鉴别这两种菌感染血清抗体的简单方法,一般的实验室都可采用。     

耶氏菌属毒力株共同血清学特性

<正> 取氏菌属细菌(鼠疫耶氏菌、假结核耶氏菌和小肠结肠炎耶氏菌)对人和动物都能引起侵袭性疾病,他们的致病性与一分子量40～48Mdal的毒力质粒有关。该质粒编码VW抗原,自凝因子和特异外膜蛋白。我们对该属细菌毒力株的共同血清学特性进行了研究,发现采用小肠结肠炎耶氏菌酶免疫斑点试验,可以把这3种细菌的毒力株和无毒力株区别开来。     

小肠结肠炎耶氏菌毒力相关基因探针的制备

本研究采用分子生物学方法从0:8型小肠结肠炎耶氏菌(Ye菌)所携带的毒力大质粒上分离、纯化了与Sereny试验相关的毒力基因2.8kb片段,经半抗原标记作为探针。菌落原位杂交结果证明2.8kb探针可与含表达VW抗原质粒的不同型Ye菌和假结核杆菌特异性杂交,与宋氏痢疾、EIEC和大肠杆菌无交叉反应。表明2.8kb探针有高度的特异性,且它与痢疾杆菌和EIEC的Sereny反应无关,在检测耶氏菌毒株方面将成为一个很有发展前景的DNA探针     

假结核耶氏菌引起胃肠炎的调查报告

我国人的假结核耶氏菌病仅有1例报告,我们在耶氏菌病调查研究中,又发现1例假结核耶氏菌Ⅵ型引起的儿童胃肠炎感染,现报告如下。 一临床资料 患儿女,3岁于1986年11月腹痛腹泻,一日数次稀便有粘液,腹部压痛。T38.2C。大便常规:红细胞0～3/HP,脓球+。取大便培养检出耶氏菌,初疑为0:13,7小肠结肠炎耶氏菌,经抗生素治     

IgA class antibodies against Yersinia enterocolitica O:3 in patients with thyroid disease

IgM, IgG and IgA class serum antibodies against Yersinia enterocolitica O:3 and O:9 and Yersinia pseudotuberculosis 1a and 3 in 41 patients with thyroid disease and 50 healthy control persons were measured by enzyme-linked immunosorbent assay (ELISA). Concentrations of antibody levels against Yersinia enterocolitica O:9 and Yercinia pseudotuberculosis 1a and 3 did not differ significantly between the patients and controls. The median value of IgA class antibody Yersinia enterocolitica O:3 was 7.5 relative units (EIU, percentage of the reference serum) in the patients with thyroid disease and 0.7 EIU in the controls (P less than 0.01; Mann-Whitney's U-test), whereas IgM and IgG class antibodies did not show this difference. IgA class serum antibodies were especially high in patients with autoimmune thyroid diseases. These findings and two case reports support the concept that there may be a causal relationship between infections with Yersinia enterocolitica O:3 and autoimmune thyroid diseases.     

我国小肠结肠炎耶氏菌毒力株的特征

本文对我国各地分离的小肠结肠炎耶氏菌的毒力特征进行测定,所有毒力株除个别外,均有VW抗原、Vi抗原和毒力因子,它们的检测结果一致。有毒株还有一条40～50MDa的质粒带,即所谓毒力质粒,但有此毒力质粒的菌株并非都是有毒株,视其有否特异性外膜蛋白(约200KDa)而定。自凝性也是检测耶氏菌毒力的一种方法,但其敏感性与特异性均不及VW抗原测定。因此测定菌株毒力,以VW抗原、Vi抗原和毒力因子测定最特异而敏感,特别后两种方法简单、易行、可靠,可在基层单位推广使用。     

小肠结肠炎耶尔森菌尿素酶基因多态性及尿素酶活性分析

目的 了解小肠结肠炎耶尔森菌尿素酶基因的多态性以及不同致病力菌株尿素酶活性之间的关联性。方法 选取43株不同致病能力的小肠结肠炎耶尔森菌用PCR扩增尿素酶基因并进行序列测定。连同4株NCBI上尿素酶基因参考序列一起进行比对和构建聚类树。同时选取8株不同致病力的小肠结肠炎耶尔森菌进行尿素酶活性实验。 结果 在低致病性小肠结肠炎耶尔森菌中,全部的18株O∶3血清型菌株的尿素酶基因7个开放读码框(ureABCEFGD)的基因序列完全一致;在全部17株O∶9血清型菌株中,尿素酶基因7个开放读码框(ureABCEFGD)的基因序列完全一致的有16株,仅1株2/O∶9(NX1997-IE419)与其他血清型菌株的尿素酶基因序列不同;高致病性的O∶8血清型小肠结肠炎耶尔森菌以及非致病菌的尿素酶基因序列与低致病性O∶3、O∶9血清型菌株的尿素酶基因序列不同且各菌株之间基因序列差异也较大。尿素酶活性实验显示不同致病能力的小肠结肠炎耶尔森菌尿素酶活性存在多样性,无明显的规律发现。结论 低致病性O∶3、O∶9血清型的小肠结肠炎耶尔森菌的尿素酶基因分别表现出高度的保守性,NX1997-IE419的尿素酶基因可能是由O∶3血清型小肠结肠炎耶尔森菌的尿素酶基因演变而来。高致病性的1B/O∶8小肠结肠炎耶尔森菌及非致病性小肠结肠炎的尿素酶基因相对不保守。尿素酶活性试验提示我们小肠结肠炎耶尔森菌的尿素酶活性大小与其致病性的强弱可能无关联。     

Plasmids of human strains of Yersinia enterocolitica: molecular relatedness and possible importance for pathogenesis

One-hundred fifteen strain of Yersinia enterocolitica were examined for plasmids and plasmid-mediated pathogenic properties. Human strains of serotypes O:3 and O:9 harbored plasmids of 46 and 44 megadaltons, respectively, with 90% homology of DNA sequences. The plasmid-mediated properties were calcium dependence, survival in human serum, conjunctivitis provocation in guinea pigs, and O agglutinogens. One strains of serotype O:8 harbored a 42-megadalton plasmid with 75% sequence homology with plasmids of serotypes O:3 and O:9. An additional plasmid-mediated property was lethality for white mice. Filter hybridization of restriction endonuclease-digested plasmid DNA indicated that a 5.6-megadalton fragment of the plasmid of serotype O:8 had virtually no sequence homology with plasmid DNA of serotypes O:3 and O:9 and therefore may be associated with the lethal factor for mice.     

Complement Killing of Yersinia enterocolitica and Retention of the Bacteria by Leucocyte Removal Filters

We report studies on the complement sensitivity of four strains of Yersinia enterocolitica , serotypes O:3, O:9, O:5,27, and O:20, isolated from blood units involved in transfusion fatalities. Complement in fresh CPD plasma killed Y. enterocolitica within 4 h at 22°C in 100% of the experiments. The bactericidal action was serotype and complement activation pathway dependent. Both classic and alternate pathways seemed to be active, but the latter to a lesser degree. When the classic pathway was blocked by chelation of Ca²⁺ no complete killing was obtained. Complement did not enhance or condition Yersinia for leucocyte filter retention. Direct removal of Yersinia by filtration was also related to serotype; all strains were reduced by filtration in heat-inactivated plasma, and all except serotype O:5,27 were reduced in Ca²⁺ -chelated plasma. Our findings may explain why plasma products and platelet concentrates are rarely involved in Yersinia sepsis related to transfusion.     

Reactive arthritis after an outbreak of Yersinia pseudotuberculosis serotype O:3 infection

OBJECTIVE: To determine the occurrence and clinical characteristics of reactive arthritis (ReA) after an outbreak of Yersinia pseudotuberculosis serotype O:3 infection. METHODS: From 15 October to 6 November 1998, a widespread outbreak of Y pseudotuberculosis serotype O:3 occurred in Finland. A questionnaire on musculoskeletal symptoms was mailed to 38 patients with infection confirmed by culture. All patients who reported joint symptoms were interviewed by phone and their medical records of outpatient visits or hospital admission because of recent joint symptoms were reviewed. RESULTS: Thirty three of 38 (87%) patients returned the questionnaire. Reactive musculoskeletal symptoms were reported by 5/33 (15%): four patients (12%) fulfilled the criteria for ReA and one additional patient had reactive enthesopathy. The patients with ReA were adults (age range 40-47 years), whereas the patient with reactive enthesopathy was a 14 year old boy. In all patients with ReA, the arthritis was polyarticular. In addition to peripheral arthritis, other musculoskeletal symptoms included sacroiliitis (one patient), pain in Achilles tendon (one patient), and heel pain (two patients). HLA-B27 was positive in all the three patients tested. In three of four patients with ReA, the duration of acute arthritis was over six months. CONCLUSION: Y pseudotuberculosis serotype O:3 infection is frequently associated with ReA and the clinical picture is severe.     

徐州地区小肠结肠炎耶尔森菌病原学初步研究

目的了解徐州地区小肠结肠炎耶尔森菌分布及病原学特征。方法对本地区分离到的菌株进行血清学分型、生化、药敏试验,并使用聚合酶链反应(PCR)技术检测毒力基因,同时对致病性血清型(O∶3和O∶9型)菌株进行脉冲场凝胶电泳分析。结果2014份标本中共检出32株(分布17个血清型)小肠结肠炎耶尔森菌,总检出率为1.59%。致病菌株占18.75%,携带ystB基因的占21.87%。32株小肠结肠炎耶尔森菌分为3个生物型别,其中1A型占78.12%,3型18.75%,2型为3.12%。本地区分离的3株O∶3血清型菌株的PFGE带型相同。结论该地区存在致病性小肠结肠炎耶尔森菌以及由此引起的腹泻疾病,尤其O∶3型菌株属于我国O∶3血清型小肠结肠炎耶尔森菌的主要克隆系,本地区应当进一步加强该菌的监测工作。