O:3型小肠结肠炎耶氏菌外膜蛋白用于布氏菌与O:9型耶氏菌感染的鉴别诊断

0:3型小肠结肠炎耶氏菌质粒编码的外膜蛋白的电泳性质及免疫原性与0:9型耶氏菌相同。利用0:3型耶氏菌外膜蛋白为抗原的免疫斑点试验或免疫印迹试验能够明确区别布氏菌与0:9型耶氏菌感染的血清学交叉反应,较通常用0:9型耶氏菌或布氏菌为抗原,更为明显。     

利用间接血凝吸收试验鉴别布氏菌和小肠结肠炎耶氏菌抗体

<正> 牛种布氏菌与O∶9血清型小肠结肠炎耶氏菌(以下简称O∶9型耶氏菌)的交叉反应已有许多学者研究。但尚未找到一种简单实用的方法,能鉴别这两种菌的交叉抗体。 前文利用凝集吸收试验鉴别布病患者血清,获得比较满意的结果。本文报告间接血凝吸收试验来鉴别这两种菌的交叉抗体。     

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。     

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。     

布鲁氏菌和其它菌属细菌感染豚鼠的皮内变态反应及血清交叉反应

动物试验进一步证实,在布氏菌和结肠炎耶尔森氏菌O:9、土拉伦菌、埃希氏大肠杆菌0:116、0:157之间具有明显的血清学交叉反应,其中Y·e O:9菌感染动物出现的交叉反应滴度高、持续时间长。此外,试验还首次证实,Y·e O:9菌、土拉伦菌和埃希氏大肠杆菌0:157致敏动物对布氏菌素可出现阳性皮肤变态反应。其中Y·e O:9菌动物的皮肤反应强度和持续时间与S型强毒布氏菌致敏动物无差别(P<0.05);土拉伦菌和大肠杆菌O:157动物的皮肤反应轻、持续时间短,48小时后即消失;大肠杆菌0:116致敏动物对布氏菌素无反应。     

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。     

12株小肠结肠炎耶尔森氏菌再生菌特性的研究

<正> 小肠结肠炎耶尔森氏菌(简称耶氏菌)是国内近年新发现的重要致病菌。本文以耶氏菌“56×”噬菌体裂解32株O∶3型菌,获得12株再生菌,频率为37.5％,这些再生菌不再与耶氏菌各型免疫血清发生     

致病性小肠结肠耶氏菌DNA多态性及其毒力表达

目的：探讨致病小肠结肠耶氏菌（简称：耶氏菌）DNA特征及其表达。方法：用聚合酶链反应（polymerase Chain Reaction;PCR）、DNA序 列分析、随机扩增多态性DNA(Randomly Amplified Polymorphic DNA;RAPD）检测耶氏菌致病基因（adherent invasion locus;ail）,并用自凝试验、刚果红试验检测其毒力表达。结果：PCR和DNA序列分析证实致病性耶氏菌O：3、O：9、O：5,27血清型含有ail致病基因,而非致病型O：4,33、O：7,8血清型不含有ail基因。此外,O：22血清型也含有ail基因与文献报告ail核苷酸序更列同源性为88.3％。O：3、O：9、O：5,27致病血清型DNA指纹图谱相类似,O：22血清型部分DNA片段与0：3血清型相似,而非致病O：4,33血清型DNA指纹图谱完全不同。毒力试验显示致病性耶氏菌全部阳性,3株O：22血清型均有少数菌落刚果试验阳性。结论：致病性耶氏菌含有ail致病基因,DNA指纹图谱相似,但表现为多态性。O：22血清型含有ail基因,DNA指纹图谱部分与O：3血清型相似,少数菌落毒力试验呈阳性反应,该血清型可能存在潜在致病性。     

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。     

宁化县家犬感染布氏菌情况调查

<正> 1990-1991年对本县城关等七个乡镇家犬作了布氏菌调查。现将调查结果报告如下。 材料与方法 一、抗原来源 B.Canis 凝集抗原(R-SAT)由中国预防医学科学院、流行病学微生物研究所制作,批号9101;免疫斑点(IDB)抗原由福建省防疫站人兽共患病研究室提供。以O:3型小肠肠炎耶氏菌(Y.ent)和牛、羊型布氏菌为外膜换取的抗原,点于同一条硝酸纤维膜(NC膜)。     

钩端螺旋体L型单克隆抗体的研制与鉴定

目的用杂交瘤单克隆抗体技术寻找鉴定临床标本中钩端螺旋体L型的方法。方法首次以从患者分离出的黄疸出血群钩端螺旋体稳定L型免疫BALB／C小鼠，取其脾细胞与骨髓瘤细胞SP2／0融合，经有限稀释法及四次克隆化。结果筛选得1株杂交瘤细胞系1B9。该单克隆抗体除与黄疸出血群56601株凝集效价为1：3200外，与其他14群14型无交叉，而同免疫原免疫获得的小鼠血清与波摩那群56008株有1：50的交叉凝集价。结论L型稳定株与原菌型有共同抗原成分，其单克隆抗体可用于特异性鉴定。     

Autoreactive T cells can be protected from tolerance induction through competition by flanking determinants for access to class II MHC

It is not clear why the N-terminal autoantigenic determinant of myelin basic protein (MBP), Ac1-9, is dominant in the B1O.PL (H-2(u)) mouse, given its weak I-A(u)-MHC binding affinity. Similarly, how do high-affinity T cells specific for this determinant avoid negative selection? Because the MBP:1-9 sequence is embryonically expressed uniquely in the context of Golli-MBP, determinants were sought within the contiguous N-terminal "Golli" region that could out-compete MBP:1-9 for MHC binding, and thereby prevent negative selection of the public response to Ac1-9, shown here to be comprised of a V beta 8.2J beta 2.7 and a V beta 8.2J beta 2.4 expansion. Specifically, we demonstrate that Ac1-9 itself can be an effective inducer of central tolerance induction; however, in the context of Golli-MBP, Ac1-9 is flanked by determinants which prevent its display to autoreactive T cells. Our data support competitive capture as a means of protecting high-affinity, autoreactive T cells from central tolerance induction.     

Molecular epidemiological study on Escherichia coli O114 by DNA analyses]

Four strains of Escherichia coli O114:non-motile (NM) were isolated from patients supposed to have endemic diarrhea in 1989. The plasmid DNA profiles and restriction fragment patterns of chromosomal DNA digested with Not I analysed by pulsed-field gel electrophoresis (PFGE) of E. coli O114:NM strains were compared with those of other 9 strains of E. coli O114 isolated elsewhere (O114:H2, 1 strain; O114:H4, 2 strains; O114:H9, 2 strains; O114:H10, 1 strain; O114:H11, 1 strain; O114:H21, 1 strain; O114:H32, 1 strain). All of E. coli O114:NM strains showed the same plasmid DNA profile and chromosomal DNA fragment patterns. The strains of E. coli O114:NM and O114:H11, and O114:H21 and O114:H32 showed the same plasmid DNA profiles, respectively. On the other hand, these strains could be differentiated by the chromosomal DNA fingerprint patterns. The chromosomal DNA fragment pattern of all E. coli O114:NM strains is completely different from those of the other 9 control strains. We suggest that chromosomal DNA fingerprinting is useful for the epidemiological study of E. coli O114 associated endemic diarrhea.     

Study on the verotoxin-producing Escherichia coli--isolation of the bacteria from deer dung

To identify the source and route of verotoxin-producing Escherichia coli (VTEC) infection in humans, we tried to isolate VTEC from fresh deer dung collected from free-range animals in two parks during the period from August 1997 to January 1998. The results are presented below. 1) VTEC were isolated from 21 of 200 deer dung samples (10.5%), consisting of 15 of 100 samples (15.0%) collected in park A and 6 of 100 samples (6.0%) collected in park B, suggesting that the incidence of VTEC isolation differs depending on location. 2) With respect to typing of verotoxin, the 21 isolated VTEC strains consisted of 10 strains (47.6%) as VT1 producer, 5 strains (23.8%) as VT2 producer, and 6 strains (28.6%) as double producer of both types. 3) With respect to serogroup of the isolated VTEC strains, 2 strains belonged to O128:H2.1 strain each belonged to the O8:H10, O128:H12, and O169:HUT groups. The remaining 16 strains failed to be identified as particular serotypes. Regarding local distribution of the serotype, in park A, 1 strain each belonged to the O128:H2, O8:H10, and O169:HUT groups. The remaining 12 strains did not clearly show particular serotypes. In park B, 2 strains belonged to O128:H2, and 4 strains failed to show particular serotypes. The remaining 1 strains showed autoagglutination. In conclusion, we isolated VTEC strains from deer that showed types of toxin and serogroups identical to those of human VTEC. Therefore, VTEC found in deer dung could well be a source of VTEC-infectious diseases in humans.     

Characterization of the histo-blood group O(2) gene and its protein product

BACKGROUND AND OBJECTIVES: This study aimed to show the full sequence and function of the O(2) allele, and investigate whether it accounts for the incompatible expression of A antigens in gastric carcinomas of blood group O persons. MATERIALS AND METHODS: By PCR, we determined the ABO genotype of group O subjects (76 gastric carcinoma patients and 165 blood donors). Two expression constructs, encoding either the putative soluble or full-length O(2) protein, were used to transfect Sf9 cells. The expression and the activity of the O(2) protein were analysed by immunohistochemistry and enzymatic assays, respectively. RESULTS: No significant difference was detectable between the O(2) allele frequency in gastric carcinoma patients (3.9%) and blood donors (4.2%). Sequencing analysis of the O(2) allele revealed an intact reading frame identical to that of A transferase except for four nucleotide substitutions. O(2)-transfected Sf9 cells and gastric carcinomas genotyped as O(1)O(2) both expressed a protein recognized by anti-A/B transferase monoclonal antibodies. In enzymatic assays, the O(2) protein failed to show measurable A transferase activity. CONCLUSION: The O(2) allele has an intact reading frame encoding a protein immunologically related to A/B transferases and enzymatically inactive. Further, our data gave no indication that the O(2) allele is related to the phenomenon of incompatible A antigen expression in gastric cancer.     

隐球菌性脑膜炎26例临床分析

目的 总结隐球菌性脑膜炎的资料,提高对隐球菌性脑膜炎的认识。方法 回顾性总结近20年（1981年10月至2001年9月）隐球菌性脑膜炎的一般资料,诊断及治疗情况。结果 共26例患者,其中男12例,女14例,年龄5－62岁,平均35．6岁,有基础疾病者16例,其中系统红斑狼疮（SLE）9例,人类免疫缺陷病毒感染或艾滋病（HIV／AIDS）4例,其他疾病3例;有明确鸽子接触史者12例;误诊结核性脑膜炎者5例,狼疮脑病者6例;墨汁染色找到隐球菌者23例（23／26）,乳胶凝集试验抗原阳性20例（20／20）。颅内压明显增高＞300mm H2O者15例,脑室扩大行侧脑室引流者9例;12例给予两性霉素B（AmpB) 5氟胞嘧啶,6例又同时加氟康唑治疗,5例AmpB 氟康唑,1例单纯应用AmpB治疗。AmpB最大用量：AmpB10.05g 脂质体两性霉素B20g,平均用量2.6g;治愈17例,好转4例,死亡或自动出院5例。同时发现近5年隐球菌性脑膜炎病例数明显增多。结论 近年来,隐球菌性脑膜炎发病率明显增高,可能和免疫抑制剂和糖皮质激素的应用及HIV／AIDS增多有关,减少病死的关键在于提高早期诊断率,治疗仍首选AmpB加5氟胞嘧啶,侧脑室引流可减少AmpB的用量,提高治愈率,缩短疗程。     

Numerical modification of the Los Angeles classification of gastroesophageal reflux disease fails to decrease observer variation

Background: We previously reported that a new endoscopic classification of gastroesophageal reflux disease, the Los Angeles classification, showed considerable observer variation depending on the experience of the endoscopist. In the present study, we evaluated some modifications of the classification to determine whether we could decrease observer variation. Methods: Fifty endoscopic photographs, each showing four images of the squamo‐columnar junction, were prospectively obtained from 50 consecutive patients with gastroesophageal reflux disease. Two groups of eight endoscopists divided by their endoscopic experience, group 1 (100–500 procedures) and group 2 (more than 500 procedures), assessed the photographs using classifications with the following modifications: (i) addition of grade O to describe healed mucosal breaks and setting grade B as more than 5 mm or 10 mm; or (ii) addition of grade O and setting grade D as 75–99% or 100% circumferential. Results: Changing the definition of grade B or grade D did not increase the kappa values for either group of observers. Conclusions: These modifications of the Los Angeles classification were unable to decrease observer variation.     

Association of ABO gene mutations resulting in a rare B subgroup

BACKGROUND AND OBJECTIVES: B subgroups are rare and the genetic analysis reported to date has been limited. MATERIALS AND METHODS: Serological and molecular investigations were performed in blood from a B-subgroup donor. RESULTS: Red cells did not react with anti-B and anti-AB reagents. However, cells absorbed anti-B. Red cells presented positive reactions with anti-H, and saliva secreted H substance. The molecular study demonstrated a B allele with the substitutions 467C>T, 646T>A, 681G>A, 771C>T, 796C>A, 803G>C, 829G>A and an O allele with the sequence of O02. CONCLUSIONS: It is probable that the presence in exon 7 of some of the O02 substitutions could have weakened the enzymatic activity of the encoded B transferase.     

Relationship between ABO and Secretor genotype with plasma levels of factor VIII and von Willebrand factor in thrombosis patients and control individuals

In contrast to earlier reports, this study examined the relationship between plasma levels of factor VIII (FVIII) and von Willebrand factor (VWF) and ABO blood group and secretor status at the genetic level in 355 patients with venous thrombosis as well as in 236 controls. ABO glycosyl transferase alleles A(1) and B were more frequent in the thrombosis collective and alleles O(1), O(2) and A(2) were more frequent in the controls. A low-risk group for venous thrombosis of individuals with genotypes O(1)O(1), O(1)O(2) and O(1)A(2) (H-antigen rich) could be distinguished from a high-risk group with genotypes A(1)A(1), A(1)B, O(1)A(1) and O(1)B (H-antigen poor). In both the thrombosis and control groups, the H-antigen rich group showed significantly lower levels of FVIII coagulant activity (FVIII:C) and VWF antigen (VWF:Ag) than the H-antigen poor group. The frequency of the different secretor genotypes in the thrombosis group was not different from that in the control group. No significant differences of FVIII:C and VWF:Ag levels were seen between SeSe, Sese and sese individuals in the thrombosis and in the control group. Thus the risk of venous thrombosis is associated with the ABO blood group genotype but not with secretor status.     

Use of antifilarial IgG4-ELISA to detect Brugia malayi infection in an endemic area of Malaysia

Summary objective   To evaluate the usefulness of antifilarial IgG4 antibody assay in detecting B. malayi infection in a filaria endemic area in Malaysia. methods   A sandwich ELISA using B. malayi soluble antigen was employed to detect antifilarial IgG4 antibodies in serum samples of 330 individuals who comprised 88 healthy individuals from nonendemic areas, 15 B. malayi microfilaraemic cases, 22 individuals with soil-transmitted helminthiases, 9 elephantiasis cases and 196 residents from a B. malayi -endemic area. An O.D. value of > 0.420 at serum dilution of 1:400 was used as the cut-off point. This cut-off point was obtained by taking the mean optical density (0.252 + 4 S.E.) of 36 negative sera which had O.D. values greater than 0.1 at serum dilution of 1: 400. results   All 15 microfilaraemic persons were positive for antifilarial IgG4 antibody. Non-endemic normals, soil-transmitted helminth infected persons and chronic elephantiasis cases were negative for antifilarial IgG4 antibody. Of the 196 individuals from the filaria endemic area, 37 (18.8%) demonstrated presence of antifilarial IgG4 antibodies; and only eight individuals (4.1%) were positive for microfilariae. All eight microfilaraemic individuals were also positive for antifilarial IgG4 antibodies. conclusion   Antifilarial IgG4-ELISA could detect 4.6 times more positive cases than the microfilaria detection method. With appropriate cut-off values that eliminate cross-reactivities, this serological tool is very useful for Brugia malayi prevalence surveys and diagnosis.