猫病毒血清学检测方法的建立及初步应用

通过摸索猫鼻气管炎病毒、嵌杯状病毒、泛白缅胞减少症病毒的培养条件,制备兔抗猫IgG．建立二种检测猫病毒抗体血清学方法即免疫酶染色法(IEA)及免疫荧光法(IFA)．用两方法检测5份猫血清标本,IEA与IFA的阳性数均为4份,结果表明IEA和IFA都适用于猫病毒抗体的检查。     

Reducing the risk of transfusion-transmitted rickettsial disease by WBC filtration, using Orientia tsutsugamushiin a model system

BACKGROUND: Careful donor screening and infectious disease marker testing have significantly reduced the incidence of transfusion-transmitted diseases and improved the safety of the blood supply. However, transfusion-transmitted diseases resulting from the use of asymptomatic yet infectious donors continue to put patients at risk. This study was undertaken to determine if third-generation WBC filters could remove Orientia tsutsugamushi-infected cells from contaminated blood. STUDY DESIGN AND METHODS: Packed RBCs were inoculated with human MNCs infected with O. tsutsugamushi at levels estimated to occur in asymptomatic infectious donors. WBC reduction was accomplished with a third-generation WBC filter. Prefiltration and postfiltration specimens were collected, serially diluted, and injected into mice to determine the infectivity of the samples. RESULTS: Mice receiving WBC-reduced packed RBCs showed no signs of illness or markers of infectivity, which suggested that a reduction of as much as 10(5) infectious rickettsiae could be achieved by filtration. CONCLUSION: The high-efficiency, third-generation, WBC-reduction filters that were tested may provide protection against the transfusion transmission of scrub typhus rickettsiae by removing from contaminated blood cells that contain intracellular bacteria.     

间接免疫荧光法检测咽拭子和尿残渣麻疹病毒抗原的诊断价值

目的探讨间接免疫荧光法(IFA)检测咽拭子和尿残渣麻疹病毒抗原对儿童麻疹病毒(MV)感染的诊断价值。方法对68例怀疑为麻疹的患儿进行咽拭子和尿残渣麻疹病毒抗原检测,分析其阳性检出率,并对部分患儿进行短期随访,分析IFA方法的临床应用价值。结果 68例患儿中有62例诊断为麻疹,咽拭子、尿残渣麻疹病毒抗原的阳性检出率分别为91.9%(57/62)和96.8%(60/62),差异无统计学意义(χ2=1.363,P0.05);62例麻疹患者中有45例诊断为典型麻疹,咽拭子、尿残渣麻疹病毒抗原的阳性检出率分别为95.6%(43/45)和97.8%(44/45),差异无统计学意义(χ2=0.345,P0.05);17例诊断为非典型麻疹,咽拭子、尿残渣麻疹病毒抗原的阳性检出率别分别为82.4%(14/17)和94.1%(16/17),差异无统计学意义(χ2=1.133,P0.05);6例诊断为其他出疹性疾病。结论 IFA法检测咽拭子和尿残渣麻疹病毒抗原是一种早期、快速、敏感的实验室方法。     

Generation of T cell responses targeting the reactive metabolite of halothane in mice

Immune-mediated adverse drug reactions (IADRs) represent a significant problem in clinical practice and drug development. Studies of the underlying mechanisms of IADRs have been hampered by the lack of animal models. Halothane causes severe allergic hepatitis with clinical features consistent with an IADR. Our ultimate goal is to develop a mouse model of halothane hepatitis. Evidence suggests that adaptive immune responses targeting liver protein adducts of the reactive metabolite (trifluoroacetyl (TFA)) play an important role in the pathogenesis. The present study demonstrated that the combination of an anti-CD40 antibody (Ab) and a Toll-like receptor (TLR) agonist served as a potent adjuvant in generating TFA-specific T cell responses in mice. Both CD4⁺ and CD8⁺ subsets of T cells were activated and the TFA-specific responses were detected not only in the spleen but also in the liver of mice immunized with mouse serum albumin adducts of TFA (TFA-MSA) plus the combined CD40/TLR agonist. Whereas all three TLR agonists examined were effective in eliciting TFA-specific immune responses in BALB/cByJ mice, only polyI:C was effective in DBA/1 mice and none of the TLR agonists could aid the generation of TFA-specific T cells in C57BL/6J mice. This result, combined with our previous finding that BALB/cByJ mice were the most susceptible to halothane-induced acute liver injury, provides the basis for employing this strain in future studies. Collectively, our data demonstrated the successful completion of a crucial first step in the development of a murine model of halothane hepatitis.     

间接免疫荧光试验在检测风疹病毒感染中的应用

目的建立检测风疹病毒(RV)抗原E1、E2、C的间接免疫荧光试验(IFA),为快速、敏感地检测临床标本中的RV提供一种重要的检测手段。方法采用荧光素标记的羊抗鼠IgG抗体,检测病毒抗原与抗风疹病毒衣壳蛋白E1、E2、C蛋白的单克隆抗体的结合物。用该系统测定中国麻疹实验室网络送检的咽拭子标本,并与病毒分离结果和临床诊断进行比较。结果IFA方法可以快速、敏感地检测临床标本中的RV。经检测咽拭子标本38份,其中28份采集于临床诊断风疹病例;6例采集于出疹患者,但诊断不明;4份采集于风疹病例的接触者。检测结果有22份风疹阳性标本。IFA的结果与病毒分离结果完全一致,但与临床诊断不一致,IFA结果的阳性数低于临床诊断风疹病例数。结论IFA方法简单可行、敏感特异,结果易于判断,是中国麻疹实验室网络中进行RV检测的重要方法之一。同时IFA方法也可以作为临床快速诊断RV感染的备选方法之一。     

实验动物弓形虫几种检测方法的应用评价研究

目的:评价检测实验动物弓形虫的2种方法。方法:应用间接免疫荧光法(IFA)、间接酶联染色法(IEA)、间接酶联免疫吸附法(ELISA)对实验动物的血清样品进行检测,以间接免疫荧光(IFA)法作为检测弓形虫的标准方法,比较后2种方法的检出率、敏感性、特异性等。结果血清经IFA、IEA、ELISA检测,分别有32.73%(18/55)、41.86(36/86)和6.34%(4/63)阳性;IFA、IEA两种方法检测血清的阳性率之间无显著性差异(P>0.05),IFA与ELISA两种方法检测血清的阳性率之间有显著性差异(P<0.05)。IEA、ELISA敏感性和特异性分别为100%与86.48%,13.33%与100%;经独立性检验,IFA与IEA两种方法存在关联性(P<0.05),IFA与ELISA两种检测方法相互独立(P>0.05)。结论:与IFA相比,IEA检测弓形虫抗体具有相似的敏感性及特异性,检出率较高,可适用于基层单位实验动物弓形虫抗体的检测;ELISA检测弓形虫抗体敏感性不高,检出率低,仍需进一步探讨和完善。     

上海市流动人员疟疾血清流行病学调查

目的 了解外来流动人员的疟疾抗体水平和相关影响因素，进而提出针对性防治措施。方法 以居住在上海市浦东、闵行2区6个镇的外来流动人员为调查对象，开展问卷调查和血清学调查，以本地户籍居民为对照。结果 共调查流动人员2732人，IFA阳性率为3．81％，按来源地流行状况分组显示，各组间疟疾IFA阳性率差异有显著性，其中来自高疟区IFA阳性率为8．10％，明显高于其它发病组（发病率〈1/万）；显示IFA阳性率与当地上报发病率基本一致。上海市本地居民IFA阳性率为1．36％与外来流动人口阳性率间差异有非常显著性；外来流动人员中阳性者主要集中在17-54岁和小学、初中学历且来沪居住时间短于5年人群。结论 来自较高疟区的流动人员输入可能对低疟区的流行带来潜在威胁，应加强对外来流动人员的疟疾监测。     

Evaluation of a recombinant antigen enzyme-linked immunosorbent assay (ELISA) in the diagnostics of antinuclear antibodies (ANA) in children with rheumatic disorders

The reliability and accuracy of ELISAs for the detection of circulating ANA in children with rheumatic diseases has recently been questioned. In this study we evaluated an allegedly superior ELISA method using recombinant antigens in a paediatric population with various rheumatic conditions and compared it to a conventional Hep-2 IFA assay. Sera from 123 children (204 blood samples) were simultaneously tested by conventional ANA immunofluorescence on Hep-2 cells (ANA-IFA) and recombinant antigen ELISA (rELISA). There were 44 children with systemic lupus erythematosus (SLE), 29 with juvenile rheumatoid arthritis (JRA), eight with mixed connective tissue disease (MCTD), eight with reactive arthritis, five with juvenile fibromyalgia syndrome, three with dermatomyositis (JDMS) and 31 with other diagnoses.  Thirty-five children (27%) had a positive Hep-2 result, which remained undetected by ELISA (P <0.002). Almost all of these children had significant IFA titres above 1:160. The major discrepancies were observed in children with JRA and SLE. There was no titre correlation between the two assays and the rELISA’s OD readings were not linear.  The ELISA using recombinant antigens was not useful for the detection of serum ANA in children with rheumatic diseases due to a high rate of false negative results. These data concur with recent reports about the lack of reliability of ELISAs using non-recombinant antigens. Received: 2 May 2001 / Accepted: 10 September 2001