甘肃省迭部和铧尖地区4种蜱自然感染莱姆病伯氏疏螺旋体的调查

目的 调查甘肃省迭部和铧尖地区4种蜱自然感染莱姆病伯氏疏螺旋体(Bb)情况,为莱姆病的防治提供科学依据。方法 2010年3月至6月,在甘肃省迭部和铧尖地区岷山北麓迭部林区（秦岭山脉）和肃南祁连山北麓铧尖林区,对4种优势蜱（森林革蜱、草原革蜱、日本血蜱和青海血蜱）自然感染莱姆病Bb进行流行病学检测。采用夹夜法,每隔10 m布夹,晚放晨收,将捕获的啮齿类动物逆毛检虫法采集寄生蜱,同时采用拖旗法采集游离蜱。对采集的4种活的成蜱,清洗消毒后解剖取其中肠内容物分别涂片,置暗视野镜下观察莱姆病Bb;对所分离到的螺旋体再用Bb单克隆和多克隆抗体鉴别试验加以证实。结果 共采集到蜱类2科8属36种,即硬蜱科6属33种,软蜱科2属3种。解剖森林革蜱、草原革蜱、日本血蜱和草原硬蜱4种201只蜱的肠道,暗视野观察出携带莱姆病Bb的阳性蜱25只,阳性率为12.44％(25/201);接种培养森林革蜱、草原革蜱和日本血蜱3种65只蜱,从12只蜱体内培养分离出莱姆病Bb,阳性率为18.46％(12/65)。结论 森林革蜱、草原革蜱和日本血蜱均有程度不同地莱姆病Bb的自然感染。     

Seronegative Lyme arthritis caused by Borrelia garinii

A case of a female patient suffering from Lyme arthritis (LA) without elevated antibody levels to Borrelia burgdorferi sensu lato is reported. Seronegative Lyme arthritis was diagnosed based on the classic clinical manifestations and DNA-detected Borrelia garinii in blood and synovial fluid of the patient, after all other possible causes of the disease had been ruled out. The disease was resistant to the first treatment with antibacterial agents. Six months after the therapy, arthritis still persisted and DNA of Borrelia garinii was repeatedly detected in the synovial fluid and the tissue of the patient. At the same time, antigens or parts of spirochaetes were detected by electron microscopy in the synovial fluid, the tissue and the blood of the patient. The patient was then repeatedly treated by antibiotics and synovectomy has been performed. Received: 28 August 2001 / Accepted: 1 January 2002     

Repeated detection ofBorrelia burgdorferi DNA in synovial fluid of a child with Lyme arthritis

Summary In the pathogenesis of late Lyme borreliosis the relative importance of the causative organism,Borrelia burgdorferi, and the immune response of the host, including autoimmune phenomena, is not yet known. We describe a 7-year-old boy with Lyme arthritis from whom two synovial fluid samples were obtained 5 months apart and up to 17 months after the first appearance of arthritis. Both synovial fluid specimens were shown to contain borrelial DNA by nested polymerase chain reaction for the amplification of portions of the genes for flagellin and OspA. Thus,Borrelia burgdorferi may persist within the joint even during late stages of the disease.     

Frequencies of Borrelia burgdorferi-reactive T lymphocytes in Lyme arthritis

Summary Using a limiting dilution system, frequencies of Borrelia burgdorferi-reactive T cells were determined in the blood and synovial fluid of four patients with chronic Lyme arthritis (LA), one patient with acrodermatitis chronica atrophicans (ACA), two patients with other inflammatory joint diseases, and two healthy individuals. B. burgdorferi-reactive precursor T cells ranged from 1/750 to 1/8 220 in case of LA and ACA patients and from 1/820 to 1/31 400 in case of controls. In vivo activated B. burgdorferi-reactive T cells were almost absent in control subjects. With one exception, they were detected in LA patients at frequencies ranging from 1/1 300 to 1/15 400. Interestingly, even after successful antibiotic therapy of LA patients, similar frequencies of in vivo activated B. burgdorferi-reactive T cells were observed in the peripheral blood, provided that low cell concentrations were used for culture. At higher cell numbers, the fraction of B. burgdorferi-reactive T cells apparently dropped, suggesting regulatory phenomena.     

一种莱姆病螺旋体real-time PCR方法的建立及其在鼠标本检测中的应用评价

目的 基于莱姆病螺旋体recA 基因,建立一种检测鼠中莱姆病螺旋体的real-time PCR方法。方法 通过GenBank分析比较莱姆病螺旋体recA 基因,选择其保守序列设计MGB探针及引物并进行方法学评估。并应用建立的real-time PCR方法和nested PCR方法对收集的123份鼠标本进行检测分析。结果 本研究建立的real-time PCR方法仅对莱姆病螺旋体检测阳性,其最小检出浓度为10¹ copies/μL。标准曲线各浓度点Ct值批内、批间平均变异系数(CV)分别为1.56%和2.30%。123份鼠标本中,real-time PCR检测59例阳性,nested PCR检测43例阳性。结论 新建立的real-time PCR方法具有快速、敏感和特异的优点,可用于鼠标本中莱姆病螺旋体的检测。     

1998-2003年河南石油勘探局新疆探区人群莱姆病监测结果分析

目的对河南石油勘探局新疆探区人群莱姆病进行监测,了解流行状况,为防治工作提供科学依据。方法采用间接免疫荧光抗体法(IFA)和酶联免疫吸附试验(ELISA),于1998-2003年对河南石油勘探局新疆探区人群莱姆病进行血清流行病学监测,结果统计学分析采用χ2检验。根据流行病学资料、临床表现和血清学检查结果进行病例诊断。结果探区人群抗莱姆病螺旋体抗体(IgG和IgM)阳性率为12.07%～14.07%,平均阳性率为13.03%(1037/7956)。不同职业人群中,莱姆病感染率以野外勘探开发组最高(16.36%),野外建设施工组次之(12.63%),后勤服务组最低(7.58%),且3组间感染率差异有显著统计学意义(χ2=101.1,P<0.01)。感染者中最小年龄6岁,最大年龄61岁,但不同年龄组感染率差异无统计学意义(χ2=7.1,P>0.01)(表2)。诊断为莱姆病者665例,患病率为8.35%。结论河南石油勘探局新疆探区人群莱姆病感染和患病率一直处于较高水平,应加强莱姆病的防治,以保护其健康。     

从全沟硬蜱分离的伯氏疏螺旋体的实验研究

本文报道了从全沟硬蜱分离的一株伯氏疏螺旋体(VL株)实验研究的结果。该株螺旋体同莱姆病螺旋体标准株在超微结构上相近,可以和高稀释度的抗伯氏疏螺旋体抗体发生间接免疫荧光反应。SDS-PAGE电泳结果显示其蛋白组成和标准株的蛋白图谱相同。热变性温度法测定其DNA的G+Cmol％含量为28.1％,和标准株的含量无明显区别,试验结果证实此株螺旋体属伯氏疏螺旋体。     

莱姆病治疗进展

莱姆病是一种由伯氏疏螺旋体所引起的,以硬蜱为主要传播媒介的自然疫源性疾病。对莱姆病的治疗主要是早期使用抗生素。莱姆病是一种全身性慢性传染病,临床表现复杂多样,侵犯多系统多器官引起损伤,根据其病程发展可分为早中晚三期,而针对不同分期、不同表现及不同发病人群的莱姆病其治疗方法又各不相同,本文将对此进行详细综述。     

Chronic Coinfections in Patients Diagnosed with Chronic Lyme Disease: A Systematic Review

PurposeOften, the controversial diagnosis of chronic Lyme disease is given to patients with prolonged, medically unexplained physical symptoms. Many such patients also are treated for chronic coinfections with Babesia, Anaplasma, or Bartonella in the absence of typical presentations, objective clinical findings, or laboratory confirmation of active infection. We have undertaken a systematic review of the literature to evaluate several aspects of this practice.MethodsFive systematic literature searches were performed using Boolean operators and the PubMed search engine.ResultsThe literature searches did not demonstrate convincing evidence of: 1) chronic anaplasmosis infection; 2) treatment-responsive symptomatic chronic babesiosis in immunocompetent persons in the absence of fever, laboratory abnormalities, and detectable parasitemia; 3) either geographically widespread or treatment-responsive symptomatic chronic infection with Babesia duncani in the absence of fever, laboratory abnormalities, and detectable parasitemia; 4) tick-borne transmission of Bartonella species; or 5) simultaneous Lyme disease and Bartonella infection.ConclusionsThe medical literature does not support the diagnosis of chronic, atypical tick-borne coinfections in patients with chronic, nonspecific illnesses.     

吉林省野鼠家畜感染伯氏疏螺旋体的检查

目的　对 1999～ 2 0 0 2年吉林省有代表性的 5个地区野鼠、家畜进行了莱姆病感染调查。方法　采用血清流行病学和病原学研究方法 ,应用间接酶联免疫吸附实验 (ELISA)检测动物血清中的IgG抗体 ,对鼠类肾脏和膀胱经研磨接种BSKⅡ培养基 ,常规 33℃培养后镜检 ,同时用直接免疫荧光抗体染色法带菌检测。结果　对野鼠、牛、马、狗、羊进行的血清学调查 ,表明其感染率分别为 :2 2 2 4 %、2 7 4 8%、16 6 7%、2 2 0 3%、16 6 6 % ;病原学培养发现 5种鼠类脏器带菌率平均分别为 :黑线姬鼠 16 35 %、黑线仓鼠 6 78%、褐家鼠 7 6 9%、小家鼠 2 2 3%、大仓鼠 6 6 7% ,从黑线姬鼠的鼠胎盘分离出一株莱姆病螺旋体。结论　实验证实莱姆病可通过野外鼠类胎盘垂直传播 ,莱姆病疫源地在吉林省广泛存在。     

Changes in the protein profile and antigenicity of different Borrelia burgdorferi strains after reintroduction to Ixodes ricinus ticks

Eight Swiss strains of Borrelia burgdorferi, with various protein profiles and the North-American strain B31 were artificially introduced into Ixodes ricinus ticks and reisolated 10 days later. All isolates were subsequently examined by SDS-PAGE analysis. Comparing initial isolates with the reisolates, we observed that 7 out of 9 strains changed their protein pattern with respect to the major proteins OspA, OspB and the 22 kDa protein after passage in the tick. The strains NE2, NE4 and NE83 with the initial phenotype of OspA and 22 kDa proteins changed to the phenotype of OspA and OspB, the strains B2 and NE202 with the initial phenotype of OspA acquired an additional protein of 22 kDa and the strain NE58 with the initial phenotype of OspA also acquired a protein of 22 kDa. Examination of these isolates by Western blot analysis demonstrated that the reaction with the monoclonal antibody H5332 and a monospecific polyclonal antibody PoAb/anti-22 kDa differed between the initial isolates and the reisolates.     

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。     

新疆博乐和沙湾、阜康市县山地莱姆病螺旋体(Borrelia burgdorferi)的分离及鉴定

1991年5月从新疆博乐市准噶尔阿拉套山山地(E81°18′,N45°13′)、沙湾县北天山西段依连哈比尔尕北坡(E85°40′,N43°55′)和阜康县北天山东段博格达山山地北坡(E88°08′,N43°55′)共采集到全沟硬蜱(Ixodes persulcatus)千余只,经做莱姆病病原体分离,共检出疏螺旋体6株。对其中5株做了单克隆抗体试验,5株疏螺旋体均与单克隆抗体H_(9724)呈阳性反应,而与H_(6831)均呈阴性反应,有3株与H_(5332)呈阳性反应、2株呈阴性反应。从而确认所分离的5株疏螺旋体为Borreliaburgdorferi疏螺旋体。首次从病原学证实上述一市二县山地存在莱姆病。     

从福建北部林区粒形硬蜱体内分离出一株莱姆病螺旋体

从福建北部已发现莱姆病病例的林区捕获一批粒形硬蜱,取其中肠接种于BSKⅡ培养基,结果分离出一株莱姆病螺旋体(Borrelia burgdorferi),从该蜱分离出莱姆病病原体在国内外属首次报告。     

新疆莱姆病的地理分布及流行情况调查

经１９９１－１９９５年对新疆维吾尔自治区内１５个县市调查，从５个县市的全沟硬蜱体检出２６株莱姆病螺旋体，血清学检查证实了１１个县市人群存在莱姆病感染，感染率为９．５％，全沟硬蜱为传播本病的主要媒介，病原体主要蛋白电泳图谱明显不同于北美菌株，调查表明莱姆病在新疆北疆山地广泛分布。     

莱姆病伯氏疏螺旋体在黑线姬鼠体内垂直传播的证实

我们于1992年在辽宁省桓仁县开展莱姆病动物宿主调查时,从黑线姬鼠的胎鼠肝中分离到1株伯氏疏螺旋体,命名为辽6。该分离株与单克隆抗体H5332、H9724反应良好,与H6831不反应(B31株均反应),分离株与国际标准株B31(分离自美国达敏硬蜱)比较,其抗原性质相似,但不完全相同。从黑线姬鼠的胎鼠中分离到伯氏疏螺旋体为世界首次报告,证明该螺旋体在小型野鼠体内可能存在经胎盘的垂直传播过程,此种传播途径在维持莱姆病自然疫源地的连续存在方面具有一定意义。     

T cell responses to polypeptide fractions of Borrelia burgdorferi in patients with Lyme arthritis

Among 6 patients with prolonged episodes of Lyme arthritis, the mean response of peripheral blood lymphocytes (PBL) to all Borrelia burgdorferi antigens (stimulation index [SI] 46) was greater than that among 5 patients with brief attacks of Lyme arthritis (SI 13; P less than 0.1), as well as that among 7 control patients with rheumatoid arthritis and among 6 normal control subjects (in both instances SI 3; P less than 0.05). In individual patients with brief episodes of Lyme arthritis, PBL had similar low levels of reactivity with the 20-kd, 31-kd, 34-kd, 41-kd, 55/58-kd, and 66-kd spirochetal polypeptides. In individual patients with prolonged arthritis, PBL usually had similar marked responsiveness to the 34-kd, 41-kd, 55/58-kd, and 66-kd polypeptides, but they had greater reactivity with the 34-kd outer surface protein B than with the 31-kd outer surface protein A (P less than 0.05). In the 2 patients tested, paired samples of synovial fluid lymphocytes and PBL had a similar pattern of reactivity, but the response was 2-100-fold greater in synovial fluid lymphocytes. We conclude that patients with prolonged Lyme arthritis have T cell responses to multiple spirochetal polypeptides.     

莱姆病的诊断与治疗进展

莱姆病是一种主要发生在温带地区经蜱叮咬传播的人兽共患自然疫源性疾病。近年来,随着蜱媒宿主范围的不断扩大,莱姆病在我国的发病率呈不断上升态势。该病系全身性疾病,可影响皮肤、关节、神经系统和心脏等。患者如得不到早期诊断和及时治疗,致残率较高,对人类健康危害甚大。鉴于莱姆病的临床表现多样,诊断困难,为了提高临床一线医护人员对莱姆病的认知,现将其临床研究进展作一综述。     

伯氏疏螺旋体外膜蛋白C基因在大肠杆菌中的表达

目的　体外表达伯氏疏螺旋体外膜蛋白Ｃ（ＯｓｐＣ） ,以制备基因工程重组抗原用于莱姆病血清学诊断研究。方法　应用聚合酶链反应技术和基因重组技术,从２ 个伯氏疏螺旋体中国分离株基因组ＤＮＡ 中扩增得到ＯｓｐＣ基因,分别克隆到表达载体ＬＫＢ２ 中,在大肠杆菌中进行表达。结果　以天然蛋白形式高效表达了ＯｓｐＣ。扫描分析显示,表达的ＯｓｐＣ 相对分子质量均在２３ ０００ 左右,表达量占菌体总蛋白的３４ ％ ～５０ ％ 。Ｗｅｓｔｅｒｎ ｂｌｏｔ 试验证明,重组ＯｓｐＣ 蛋白可与莱姆病患者血清发生特异性反应。结论　重组ＯｓｐＣ 的成功表达为制备新型莱姆病早期诊断试剂奠定了基础