伯氏疏螺旋体表膜蛋白C（OspC）的研究和应用

伯氏疏螺旋体（Borrell’abundorferi）是蚂媒疫源性疾病一莱姆病的病原体，侵人机体后可造成多器官系统受损，患者早期可出现慢性游走性红斑、流感样症状，晚期可导致心肌炎、关节炎及神系统严重病变，重者可危及生命。自1975年首例患者确证以来，莱姆病已成为美国最为常见的蚂传播性疾病，随着欧洲亚洲的中国、日本及其它一些国家病例报道日益增多，莱姆病已作为世界性卫生课题受到越来越多的重视’‘·”。如何做到早期诊断及预防一直是众多学者研究的重点。由于莱姆病缺乏特异性临床表现，诊断主要依据实验室检测，其中血清学检测以其…     

莱姆病的研究近况

莱姆病是一种新发现的、由蜱传伯氏疏螺旋体（Ｂｏｒｅｌｉａｂｕｒｇｄｏｒｆｅｒｉ）感染引起的人兽共患疾病，临床上主要表现为皮肤、心脏、神经及关节等多器官的损害。自１９８２年Ｂｕｒｇｄｏｒｆｅｒ及其同事发现和分离出莱姆病的致病因子以来，莱姆病在流行病学、...     

莱姆病螺旋体鞭毛平截性蛋白的表达及其诊断潜力研究

目的原核表达伯氏疏螺旋体鞭毛蛋白Flagellin A基因特异性区段,获得重组鞭毛蛋白平截性蛋白作为诊断抗原,建立间接ELISA方法用于动物莱姆病的诊断。方法 PCR扩增获取伯氏疏螺旋体鞭毛蛋白基因的同源性较低的第394-798bp区段,构建重组质粒pGEX-4T-1/tFlaA,构建好的表达质粒转化到大肠杆菌BL21（DE3）中进行表达,并纯化重组蛋白,用纯化的表达蛋白作为莱姆病诊断的抗原,用于ELISA检测实验感染小鼠莱姆病。结果成功构建莱姆病螺旋体鞭毛平截性蛋白的表达载体,重组蛋白在宿主菌内高效、稳定表达,重组平截性蛋白显示了可作为ELISA诊断的抗原用于莱姆病的诊断价值。结论纯化的伯氏疏螺旋体鞭毛蛋白可作为莱姆病ELISA诊断抗原用于莱姆病的诊断,为莱姆病快速诊断试剂盒的开发打下基础。     

肠道病毒性脑膜炎基因诊断的研究

肠道病毒（ＥＶＳ）感染是无菌性脑膜炎和脑炎常见的病因之一，由于其临床表现及脑脊液（ＣＳＦ）检查无特异性，故临床难与其它细菌性或病毒性感染相鉴别。本文用逆转录聚合酶链反应（ＲＴ－ＰＣＲ）检测了临床诊断的２４例病毒性脑膜炎及２０例其它神经疾病（ＯＮＤ）对照组患者的ＣＳＦ中ＥＶｓ－ＲＮＡ。所用的引物选自ＥＶｓ基因组中高度保守的５＇－非编码区（５＇－ＮＣＲ），这些引物已被证明能够检测出ＥＶｓ广泛的型别。结果在２４例病毒性脑膜炎患者ＣＳＦ中检出ＥＶｓ－ＲＮＡ阳性者７例，而对照组全部阴性。提示经ＲＴ－ＰＣＲ的早期诊断有助于对ＥＶｓ脑膜炎采取早期特异的抗病毒治疗。     

莱姆病螺旋体感染与精神疾病发生关系初探

为了解莱姆病螺旋体感染后期是否引起精神障碍，作者采用间接免疫荧光抗体试验，对内蒙古大兴安岭林区１３８例各类精神疾病患者及９０例健康对照者进行了血清抗莱姆病螺旋体抗体检查和分析，现报道如下。１资料和方法入组标准符合中国精神疾病分类与诊断标准第二版，诊断...     

伯氏疏螺旋体感染性认知障碍的研究现状

莱姆病是由伯氏疏螺旋体(BB)感染人体,引起全身多器官病变的一种螺旋体虫媒传染病.在美国和欧洲莱姆病是最常见的通过节肢动物传播的一种疾病,亚洲也有发现.每年大约有20 000例新发病例.能引起莱姆病的BB有狭义伯氏疏螺旋体(Borrelia burgdorferi sensust ricto)、伽氏疏螺旋体、阿佛西尼疏螺旋体.当神经系统的表现比较突出时,称之为神经莱姆病.累及神经系统的螺旋体基因型主要是是伽氏疏螺旋体.20多年前发现了第一例莱姆病性痴呆,至今出现认知障碍的患者越来越多.神经莱姆病患者在治疗前大约55％出现认知障碍,10％的患者表现有轻度痴呆,经过6个月的治疗之后有15％的患者有轻度痴呆.本文就神经莱姆病的流行病学、病因与发病机制、病理、临床表现、诊断、治疗与预后等做一综述.     

散发性病毒性脑膜炎135例病原学分析

散发性病毒性脑膜炎１３５例病原学分析邱杰文，谢若男，刘卫东，袁广卿，吴胤荣，廖传红病毒性脑膜炎（病毒脑）是由病毒引起的无菌性脑膜炎，为了解近年来病毒脑病原分布的情况，我们对汕头市１３５例临床诊断为病毒脑的散发性患者进行病毒分离及血清学检测，现报告如下...     

美国《Emerging Infectious Diseases》2023年第6期有关人兽共患病论文摘译

<正>P1091神经莱姆病后持续症状与血液中IFN-α水平升高的相关性//Sergio A.Hernández,Katarina Ogrinc,Mi?a Korva，等神经莱姆病（LNB）患者尽管接受抗生素药物治疗，仍可能会经历持续的症状。本文对79例LNB患者的血清和脑脊液（CSF）中20种免疫介质进行为期1年的检测，来验证这些症状是否由适应性免疫反应异常引起。在研究初期，大多数介质高度集中在CSF感染的部位。     

ELISPOT检测在结核性脑膜炎诊断中的研究

目的利用酶联免疫斑点法(ELISPOT)检测脑脊液中结核抗原特异性的INF-γ分泌细胞数量,研究其在结核性脑膜炎诊断中的价值。方法对104例结核性脑膜炎患者和40例非结核性脑膜炎患者进行脑脊液与血的ELISPOT检测,并与脑脊液抗酸染色涂片、结核菌培养、荧光定量PCR、结核抗体检测进行敏感度和特异性的比较。结果脑脊液ELISPOT检测的敏感度为67.3%、特异性为89.2%,PV+值94.3%优于其他检测方法,差别具有统计学意义(P0.05),脑脊液ELISPOT检测具有较好的诊断价值(Kappa=0.856,P0.001)。结论脑脊液ELISPOT检测对结核性脑膜炎诊断的敏感度及特异度高,有望成为快速诊断结核性脑膜炎的一种方法。     

结核杆菌硬脂酸测定对脑脊液菌阴结核性脑膜炎诊断价值的探讨

采用毛细管柱气相色谱技术分离出结核杆菌特征组份并鉴定为结核杆菌硬脂酸。对５９例脑脊液菌阴结核性脑膜炎患者与１２２名非结核性脑膜炎对照组进行分析。实验结果显示结核性脑膜炎组检出结核杆菌硬脂酸４４例，对照组中检出５例，敏感性及特异性分别为７４．６％和９５．９％。表明结核杆菌硬脂酸的检测有助于菌阴结核性脑膜炎的早期诊断。     

Evaluation of the intrathecal antibody response to Borrelia burgdorferi as a diagnostic test for Lyme neuroborreliosis

The intrathecal antibody response to Borrelia burgdorferi was evaluated in American and West German patients with Lyme neuroborreliosis. By an antibody capture enzyme immunoassay, 12 (92%) of 13 patients from the USA with Lyme meningitis were found to have intrathecal antibody production to B. burgdorferi, usually of multiple isotypes, most commonly IgA. Of 12 patients with putative late central nervous system manifestations of Lyme disease, 5 (42%) had local production of IgG or IgA spirochetal antibody, but cerebrospinal fluid (CSF) abnormalities could not be demonstrated in 6 patients with late peripheral nervous system manifestations of the disorder. Compared with American patients, 30 European patients with neuroborreliosis had significantly higher CSF:serum ratios of specific antibody both early and late in the illness. Intrathecal antibody determinations are the most specific diagnostic test currently available for Lyme neuroborreliosis, but local antibody production in CSF is an inconsistent finding in American patients with late neurologic manifestations of the disorder.     

The T-cell proliferative assay in the diagnosis of Lyme disease

OBJECTIVE: To determine the sensitivity and specificity of the T-cell proliferative assay as a diagnostic test in Lyme disease. DESIGN: Cross-sectional study of patients with Lyme arthritis or chronic neuroborreliosis who had a history of erythema migrans, positive antibody responses to Borrelia burgdorferi by enzyme-linked immunosorbent assay (ELISA), or both; patients with other diseases; and healthy subjects. SETTING: Diagnostic Lyme disease clinic in a university hospital. PATIENTS: Forty-two of the 67 patients with active Lyme arthritis or chronic neuroborreliosis who were seen during the study period; 16 patients with inactive late Lyme disease; 77 patients with other rheumatologic or neurologic diseases; 9 workers from the Borrelia laboratory; and 9 healthy subjects. MEASUREMENTS AND MAIN RESULTS: Nineteen of 42 patients with Lyme arthritis or chronic neuroborreliosis and 4 of 77 patients with other diseases had positive T-cell proliferative responses to B. burgdorferi antigens. The sensitivity of the proliferative assay was 45% (95% Cl, 30% to 60%) and the specificity was 95% (95% Cl, 87% to 99%). Twelve of 27 patients with active Lyme arthritis, 7 of 15 patients with chronic neuroborreliosis, 4 of 16 patients with inactive Lyme disease, 4 of 9 healthy Borrelia laboratory workers, and 0 of 9 healthy subjects had positive responses. Three of five patients with Lyme disease who had negative or indeterminant antibody responses by ELISA had positive T-cell proliferative responses. CONCLUSION: The T-cell proliferative assay may be a helpful diagnostic test in the small subset of patients with late Lyme disease who have negative or indeterminant antibody responses by ELISA.     

Detection of multiple reactive protein species by immunoblotting after recombinant outer surface protein A lyme disease vaccination.

Laboratory confirmation of the diagnosis of Lyme disease is based on the detection of an immune response to Borrelia burgdorferi. The serodiagnosis of B. burgdorferi infection is complex and may be further confounded by the immune response to the recombinant outer surface protein A (OspA) Lyme disease vaccine. To describe how the serological response to the recombinant OspA Lyme disease vaccine affects testing for antibody to B. burgdorferi, 240 specimens from 80 study subjects were obtained at defined intervals after recombinant OspA Lyme disease vaccination. Samples were tested by indirect enzyme-linked immunosorbent assay (ELISA), antibody capture enzyme immunoassay (EIA), and Western blotting (WB). After recombinant OspA Lyme disease vaccination, ELISA for 98% of the study subjects revealed reactivity. WB with use of OspA-containing B. burgdorferi strains as sources of antigens demonstrated multiple bands. Results of testing with a US Food and Drug Administration-approved WB kit showed homogeneous reactivity in the molecular weight region >30 kDa. Testing with OspA-free strains completely eliminated all vaccine-associated reactivity by both antibody capture EIA and WB.     

Concomitant infection with tick-borne encephalitis virus andBorrelia burgdorferi sensu lato in patients with acute meningitis or meningoencephalitis

Summary From September 1992 to August 1993, 338 patients over the age of 15 years presented to the Department of Infectious Diseases, University Medical Centre Ljubljana, with acute lymphocytic meningitis. In 89 of these patients (26.3%) serum IgM and IgG antibodies against tick-borne encephalitis (TBE) virus were detected, and in 59 patients (17.5%) a borrelial etiology of disease was demonstrated by one or more of the following: presence of intrathecal antibody production, seroconversion to borrelial antigens, presence of erythema migrans, and/or isolation ofBorrelia burgdorferi sensu lato from skin or cerebrospinal fluid. Of the 148 patients who fulfilled criteria for TBE or borrelial infection, concomitant infection with TBE virus andB. burgdorferi sensu lato was demonstrated in 12 patients (3.6% of all patients presenting with acute lymphocytic meningitis). In the majority of patients with concomitant infection the clinical features at presentation were characteristic of, or consistent with, TBE. In addition, during follow-up studies, eight of the 12 patients subsequently developed signs and symptoms compatible with minor and/or major manifestations of Lyme borreliosis. Six patients were diagnosed with neuroborreliosis based on signs or symptoms and/or laboratory tests. These findings show that in patients with acute lymphocytic meningitis or meningoencephalitis, originating in TBE and Lyme borreliosis endemic regions, the possibility of concomitant infection should be considered.     

Antigenic conservation of an immunodominant invariable region of the VlsE lipoprotein among European pathogenic genospecies of Borrelia burgdorferi SL

Lyme disease is caused by genetically divergent spirochetes, including 3 pathogenic genospecies: Borrelia burgdorferi sensu stricto, B. garinii, and B. afzelii. Serodiagnosis is complicated by this genetic diversity. A synthetic peptide (C(6)), based on the 26-mer invariable region (IR(6)) of the variable surface antigen of B. burgdorferi (VlsE), was used as ELISA antigen, to test serum samples collected from mice experimentally infected with the 3 genospecies and from European patients with Lyme disease. Regardless of the infecting strains, mice produced a strong antibody response to C(6), which indicates that IR(6) is antigenically conserved among the pathogenic genospecies. Twenty of 23 patients with culture-confirmed erythema migrans had a detectable antibody response to C(6). A sensitivity of 95.2% was achieved, with serum samples collected from patients with well-defined acrodermatitis chronica atrophicans. Fourteen of 20 patients with symptoms of late Lyme disease also had a positive anti-IR(6) ELISA. Thus, it is possible that C(6) may be used to serodiagnose Lyme disease universally.     

Enzyme-linked immunosorbent assays for Lyme disease: reactivity of subunits of Borrelia burgdorferi

We prepared fractions of Borrelia burgdorferi, the etiologic agent of Lyme disease, from cultured spirochetes and used them as antigen in an enzyme-linked immunosorbent assay (ELISA) for IgG antibody. Polystyrene plates coated with an extract containing major proteins with apparent molecular masses of 34, 39, 59, and 68 kilodaltons had comparable sensitivity but greater specificity than plates coated with whole cells. Of the 33 serum specimens from individuals with Lyme disease that reacted with whole cells of B. burgdorferi in the class-specific ELISA, 30 (91%) remained positive when this extract was used. Cross-reactivity was minimal with antibody to treponemes. Use of subunit antigens may improve serological diagnosis of Lyme disease.     

No evidence to implicate Borrelia burgdorferi in the pathogenesis of dilated cardiomyopathy in the United Kingdom.

OBJECTIVE--To determine whether Borrelia burgdorferi is implicated in the pathogenesis of dilated cardiomyopathy in the United Kingdom. DESIGN--A controlled prospective study. Patients' notes were reviewed for evidence of Lyme disease and serum samples were tested by enzyme linked immunoadsorbent assay (ELISA) for antibodies to B burgdorferi. Samples with raised antibody concentrations were subsequently analysed by immunoblotting to determine their antibody binding specificity. SETTING--Tertiary referral centre. PATIENTS--97 consecutive patients with dilated cardiomyopathy diagnosed according to World Health Organisation criteria were studied. Serum samples were taken from two matched control groups. The first group (n = 38) was age, sex, and geographically matched. The second control group (n = 39) was environmentally matched and consisted of members of the patients' own households. MAIN OUTCOME MEASURES--Clinical evidence of Lyme disease. Presence of raised antibody concentrations to B burgdorferi. RESULTS--No patients had a previous illness compatible with Lyme disease. Analysis of the ELISA data showed eight of 97 patients with dilated cardiomyopathy (8.2%) and two of 77 controls (3.9%) had raised antibody concentrations. Immunoblot analysis, however, did not show binding patterns consistent with the presence of IgG specific for B burgdorferi in any of these samples. CONCLUSIONS--There was no clinical or serological evidence to implicate B burgdorferi in the pathogenesis of idiopathic dilated cardiomyopathy in the United Kingdom. In the absence of specific symptoms or likely exposure to B burgdorferi routine serological testing for Lyme disease in this group of patients is not recommended. Furthermore, raised antibodies to B burgdorferi are not diagnostic of active infection and ELISA results should be interpreted with caution unless specific B burgdorferi antibody bands have been found by immunoblot analysis.     

Antibody response to IR6, a conserved immunodominant region of the VlsE lipoprotein, wanes rapidly after antibiotic treatment of Borrelia burgdorferi infection in experimental animals and in humans

Invariable region (IR)(6), an immunodominant conserved region of VlsE, the antigenic variation protein of Borrelia burgdorferi, is currently used for the serologic diagnosis of Lyme disease in humans and canines. A longitudinal assessment of anti-IR(6) antibody levels in B. burgdorferi-infected rhesus monkeys revealed that this level diminished sharply after antibiotic treatment (within 25 weeks). In contrast, antibody levels to P39 and to whole-cell antigen extracts of B. burgdorferi either remained unchanged or diminished less. A longitudinal analysis in dogs yielded similar results. In humans, the anti-IR(6) antibody titer diminished by a factor of > or =4 in successfully treated patients and by a factor of <4 in treatment-resistant patients. This result suggests that the quantification of anti-IR(6) antibody titer as a function of time should be investigated further as a test to assess response to Lyme disease therapy or to determine whether a B. burgdorferi infection has been eliminated.     

Lyme borreliosis

Lyme borreliosis is a multi-organ infection caused by spirochetes of the Borrelia burgdorferi sensu lato group with its species B burgdorferi sensu stricto, Borrelia garinii, and Borrelia afzelii, which are transmitted by ticks of the species Ixodes. Laboratory testing of Lyme borreliosis includes culture, antibody detection using ELISA with whole extracts or recombinant chimeric borrelia proteins, immunoblot, and PCR with different levels of sensitivity and specificity for each test. Common skin manifestations of Lyme borreliosis include erythema migrans, lymphocytoma, and acrodermatitis chronica atrophicans. The last two conditions are usually caused by B garinii and B afzelii, respectively, which are seen more frequently in Europe than in America. Late extracutaneous manifestations of Lyme borreliosis are characterised by carditis, neuroborreliosis, and arthritis. We present evidence-based treatment recommendations for Lyme borreliosis and review the prevention of Lyme borreliosis, including the Lyme vaccines.     

中国莱姆病螺旋体特异性单克隆抗体的制备及初步鉴定

目的制备特异的莱姆病螺旋体单克隆抗体,为我国莱姆病的诊断和莱姆病螺旋体菌株鉴定提供基础。方法以中国莱姆病螺旋体伽氏疏螺旋体(Borreliagarinii)的代表菌株PD91的全菌蛋白为抗原,免疫BALB/c小鼠,取脾细胞与骨髓瘤细胞SP2/0融合,用间接酶联免疫吸附试验(ELISA)和蛋白免疫印迹方法(WB)筛选,并经过2次或3次克隆,以获得单克隆抗体。结果共制备出10株单克隆抗体,经鉴定为3种,分别针对中国莱姆病螺旋体的外膜蛋白OspA(4株)、OspB(3株)和OspC(3株)。结论成功制备出3种抗莱姆病螺旋体不同蛋白的单克隆抗体,可用于我国莱姆病的病原诊断和莱姆病螺旋体菌株鉴定。