酶联免疫吸附试验(ELISA)在流行性乙型脑炎血清学诊断上的应用

本文介绍了用ELISA检测225份乙脑患者血清标本并与CFR进行了比较。ELISA的阳性率高于CFR,乙脑病毒抗原与其它传染病阳性血清和正常人血清不发生特异性反应,并且重复性好。由于ELISA方法敏感、特异、简便,所用试剂容易制备或获得,结果读取客观,不仅可以用于乙脑的临床实验诊断,而且也适用于流行病学调查研究工作。     

免疫印迹法在献血员梅毒抗体筛查中的应用

目的研究梅毒螺旋体免疫印迹试验(TP-WB)在献血员梅毒抗体筛查中的应用价值。方法使用梅毒标准血清盘,分析2种TP-ELISA试剂和1种TPPA试剂检测的准确性及不足;使用WB法对无偿献血筛查的279份TP-ELISA试剂阳性标本进一步检测,分析ELISA阳性结果与WB结果的相关性及WB阳性条带的分布特征。结果在40份以TP-WB结果为金标准的血清盘中,XC和WT两公司的TP-ELISA试剂的检测灵敏度均为100.00%,高于TPPA的88.46%,但两公司试剂检测的特异性分别为92.86%与85.71%,均低于TPPA的100.00%;在筛查的279份TP-ELISA法检测阳性标本中,WB确认阳性216份,阳性率为77.42%;其中包括S/CO值5且两种ELISA试剂检测同时阳性的标本205份,其WB确认试验阳性率为100.00%,占216份WB确认阳性标本数的99.91%。有序Logistic回归分析显示,ELISA法S/CO值5和双试剂阳性,与WB检测阳性分别存在统计关联(P0.01);216份TP-WB阳性标本WB条带分布分析,其中TP17条带与梅毒抗体Ig M阳性、TP15条带与Ig G及Ig G+Ig M阳性分别存在统计关联(P均0.01)。结论 ELISA法双试剂阳性且S/CO值5,基本可确诊为梅毒,WB试验条带阳性结果对献血者感染状况的判断有一定的指导意义。     

西双版纳州2007年-2010年流行性乙型脑炎疑似病例IgM抗体检测结果分析

目的:通过实验室检测结果,了解西双版纳州流行性乙型脑炎(乙脑)发病情况,为乙脑诊治和制定防制策略提供依据。方法:用ELISA法对乙脑疑似病例标本进行IgM抗体检测,并根据检测结果和流行病学资料进行分析。结果:2007年-2010年共检测乙脑疑似病例标本240例,阳性149例,阳性率为62.08%;4年的阳性率依次为90.00%,57.50%,36.54%6,2.50%;阳性病例年龄最小6个月,最大72岁,以学生和散居儿童为主,占65.10%;流行季节为5月-8月份,发病呈单峰分布,高峰出现在6月份,占54.36%;男女性别比为1.61∶1;阳性病例中95.97%无接种史和疫苗接种史不详。结论:西双版纳州存在乙脑高流行的自然环境,应采取综合性防制措施,有效地控制乙脑发病率。     

探讨酶联免疫吸附试验检测乙肝核心抗体假阴性的问题

目的:了解用酶联免疫吸附试验(ELISA)检测乙肝二对半核心抗体(抗-HBc)的假阴性。方法:留取用ELISA检测HBsAg阳性、HBeAg阳性、抗-HBc阴性血清标本201份,分别用美国雅培AXSYM免疫发光仪微粒子酶免疫技术(MEIA)定量检测抗HBc、ELISA进行原倍和生理盐水15倍稀释测定。结果:MEIA法定量检测抗-HBc阳性166份,阳性检出率为82. 6%;阴性(S/Co>1 0)35份,与ELISA一步法相比阴性符合率为19 .9%,生理盐水15倍稀释阳性33份,阳性检出率为19 4%,原倍血清100% 阳性。结论:MEIA法定量检测的阳性率远远高于ELISA法检测, 15倍稀释测定结果阳性率比30倍稀释有所提高,原倍血清仅作为流行病学调查结果。     

ELISA方法检测乙肝表面抗原假阳性的分析

1材料标本:2005年3月至2006年6月经ELISA方法检测为阳性的病人标本共63例。1.1仪器及试剂方法一:ELISA,仪器:三科酶标仪,试剂:科华表面抗原ELISA试剂。方法二:电化学发光法,仪器:强生ECI电化学发光仪,试剂:强生电化学发光仪配套试剂。2结果结果判定:ELISA方法S/Co>1.0。电化学发光法S/Co>1.0。见表1。表1两种方法检测结果比较方法HBSAG阳性阳性率酶联免疫吸附试验63100%电化学发光法5790.4%从上表可知,用电化学发光法检测ELISA阳性的标本有90.4%的阳性率,其中由ELISA形成假阳性率有9.6%,造成这部分假阳性率的原因主要有:2.…     

2种登革热抗体检测试剂应用评价

[目的]探讨酶联免疫吸咐试验(EIJSA)、快速免疫层析法(ICT)检测试剂检测登革病毒(DV)抗体的敏感性、特异性及优缺点。[方法]用ELISA及ICT试剂同时检测来自登革热疫区入境人员血清标本的DV-IgM、IgG抗体,比较检测结果。[结果]用ELISA检测91份来自登革热疫区但无登革热体征的入境人员血清DV-IgG抗体,阳性59份,阳性率64．8％；对59份DV-IgG抗体阳性标本采用ICT法检测DV-IgM、IgG抗体,结果检出4份DV-IgM阳性,未检出DV-IgG阳性。ICT法检测181份血清标本DV-IgM、IgG抗体,检出12份DV-IgG阳性,阳性率6．63％；2份DV-IgM阳性,阳性率1．10％；其中44份血清标本采用ICT试剂不同批号检测DV-IgM、IgG抗体,批号为10014c- 12405的检出DV-IgM阳性7份,未检出DV—103阳性；批号为10014c一24505的检出4份DV-IgG阳性,1份DV-IgM阳性。[结论]2种试剂的敏感性和特异性及不同批次之间的稳定性有待进一步论证。     

健康人群与猪感染乙型脑炎病毒状况分析

目的通过调查健康人群以及乙脑动物宿主猪乙脑抗体水平变化,为有效预防乙脑提供科学依据。方法采用纵横结合研究,在观察点饲养未经历乙脑流行期的仔猪12头,按旬对每只猪采集血清,每份血液采集4ml,应用间接法ELISA检测乙脑抗体;在观察点所在村,按整群抽样的方法,在乙脑流行季节前后（4月、11月）对调查点健康人群各采血1次,应用间接法ELISA检测乙脑抗体。结果共采集猪血清标本221份,乙脑抗体阳性150份,抗体阳性率为67．87％。在乙脑流行季节前后分别采集健康人群血清标本300份、294份,乙脑抗体阳性率分别为39．33％、42．86％。结论幼猪出生后无免疫力,受蚊子叮咬后几乎100％受感染,病毒血症期长,是乙脑的主要传染源;健康人群在乙脑流行季节后乙脑抗体阳性率有所升高,但总体乙脑抗体阳性率较低,要警惕乙脑流行。建议提高乙脑疫苗免疫接种率;对猪只进行乙脑免疫;避免人猪间乙脑的相互传播。     

2010年深圳市南山区健康人群乙脑抗体水平监测

目的为了解深圳市南山区健康人群乙脑抗体水平,评价其流行危险度。方法随机抽取南山区0岁～、5岁～、10岁～、15岁～、20岁～、30岁～、40岁～等年龄组各80～90人,采集血清标本用ELISA法检测抗体。结果检测南山区健康人群血清631份,乙脑IgG抗体阳性218份,总阳性率34.55%。不同年龄组间抗体阳性率有差异,20岁以上成人组乙脑抗体阳性率较低;男女性别的抗体阳性率差异无统计学意义。结论南山区健康人群乙脑免疫水平较低,一旦有输入性病例,暴发和流行的危险度较大。     

化学发光免疫分析法与酶联免疫法检测艾滋病抗体的对照研究

目的对照研究化学发光免疫分析法(CLIA)与酶联免疫法(ELISA)检测艾滋病(HIV)抗体的效果,以便探究CLIA检测HIV抗体的可行性。方法于2013年2月至2015年3月随机抽查我市3 542例艾滋病高危人员,采集血清,分别采用CLIA与ELISA法检测血清中的HIV抗体,并采用蛋白印迹检测法对检测结果进行确证。结果 ELISA法检测显示血清标本为阳性的56份,阳性率为1.58%,而CLIA检测阳性61份,阳性率为1.72%;3 542份血清标本中,11份标本ELISA法与CLIA法检测结果不一致;CLIA法的阳性预测值为96.72%,检测灵敏度为98.33%,特异度为99.94%;ELISA法的阳性预测值为96.43%,检测灵敏度为90.00%,特异度为99.94%;ELISA法与CLIA法在阳性预测值、检测灵敏度、特异度等指标上无显著的差异性(P>0.05)。结论 CLIA的灵敏度、特异度、准确性高,可用于HIV抗体检测。     

两种方法检测575例献血者血清梅毒抗体的比较分析

目的 探讨梅毒抗体的有效检验方法。方法 采用酶联免疫吸附试验（ELISA）和梅毒螺旋体特异性抗体明胶凝集试验（TPPA）对575份献血者血清标本进行检测比较。结果 对于75份梅毒阳性血清,TP-ELISA检出阳性74份、TPPA检出阳性73份,两法的阳性检出率分别为98.7%和97.3%。对于500份非梅毒患者血清,TP-ELISA检出1份阳性,TPPA均为阴性,后经临床及实验确认,该例为假阳性,TP-ELISA的假阳性率为0.2%。结论 ELISA和TPPA结果无显著性差异（P〉O.05）。ELISA试剂成本低,操作方便,是大批量样本筛查的理想方法,应作为目前临床、血站诊断和筛查血清梅毒特异性抗体的首选。     

浙江省蚊媒携带流行性乙型脑炎病毒的分布特点

目的 了解浙江部分地区蚊媒及猪携带流行性乙型脑炎(乙脑)病毒情况以及浙江省存在的乙脑病毒基因型别.方法 于2007年5月至10月在浙江省仙居、龙泉和慈溪3个县人房及猪舍采集蚊虫标本和猪血清,共采集10 662只蚊虫和204份猪血清,蚊虫标本中以淡色库蚊和中华按蚊为主.利用病毒分离和荧光定量逆转录聚合酶链反应(RT-PCR)方法检测蚊虫携带乙脑病毒,应用酶联接免疫吸附剂测定(ELISA)方法对猪血清进行抗体检测.应用荧光定量RT-PCR方法对细胞分离株进行病毒鉴定;利用RT-PCR方法扩增新分离乙脑病毒PrM基因,并对细胞分离到的病毒进行基因分型.结果 在蚊虫标本中检出7批乙脑病毒核酸阳性样本,并分离出3株病毒,经鉴定3株病毒均为乙脑病毒,基因分析表明3株病毒均属于基因Ⅰ型乙脑病毒.猪血清有121份阳性,总阳性率为59.3%.6月份有50%以上的猪血清中乙脑病毒抗体呈阳性.结论 浙江省部分地区存在着乙脑的传播媒介,在蚊媒和猪中携带有乙脑病毒;采集的蚊虫标本中分离到3株病毒,经鉴定为基因Ⅰ型乙脑病毒,是近年来在浙江省首次分离到的基因Ⅰ型乙脑病毒.     

合肥市2008～2009年流行性乙型脑炎疑似病例血清流行病学分析

[目的]分析合肥市2008～2009年流行性乙型脑炎实验室检测结果,为预防控制乙脑等疾病提供科学依据。[方法]收集合肥市2008～2009年的乙型脑炎疑似病例526例,用ELISA法检测乙型脑炎特异性IgM抗体。[结果]共检出乙型脑炎IgM抗体阳性病例132例,年龄以1～12岁儿童居多,占总数的87.88%,男性明显高于女性;发病时间全部集中在7～9月份。[结论]除强化主动监测,加强小年龄组儿童的预防接种外,必须重视实验室监测,增强检测能力,以提高诊断的准确性。     

Serodiagnosis of Japanese encephalitis among Nepalese patients by the particle agglutination assay

Japanese encephalitis (JE) is a serious health problem in the southwestern region of Nepal. Serological diagnostic kits for routine diagnostic use in this region have not been available. This study was performed to examine if the particle agglutination (PA) assay for Japanese encephalitis virus (JEV) IgM could be applicable to the samples collected in Nepal and also to evaluate the accuracy of clinical diagnosis of JE. One hundred and ninety-three blood samples were collected from the patients clinically diagnosed with JE or other infectious diseases in the JE-endemic, southwestern region of Nepal, in 2000. The PA assay was performed on these 193 serum samples and the results were compared with those by IgM-capture ELISA. Eighty-six samples were IgM-positive by the PA assay, and 71 of 86 were also positive by IgM-capture ELISA (sensitivity, 99%; specificity, 88%; positive predictive value, 0.82; negative predictive value, 0.99). These results suggest that the PA assay is a simple, reliable and useful diagnostic test to support clinical diagnosis in rural hospitals of Asia including Nepal.     

河北省健康人群流行性乙型脑炎抗体水平调查

目的了解河北省流行性乙型脑炎健康人群抗体水平。方法在河北省4个市采用横断面研究,按整群抽样的方法,分6个年龄组,每个年龄组20人份,采集血清标本。应用间接法ELISA检测乙脑IgG抗体。结果河北省健康人群抗体阳性率为40.63%。不同年龄组人群乙脑抗体阳性率差异无统计学意义,不同地区间乙脑抗体阳性率差异有统计学意义。结论河北省健康人群乙脑抗体阳性率较低,特别是个别地区应警惕乙脑流行。要进一步抓好乙脑疫苗免疫接种工作,提高疫苗接种率和接种质量,提高人群免疫水平。     

Antibody to the nonstructural protein NS1 of Japanese encephalitis virus: potential application of mAb-based indirect ELISA to differentiate infection from vaccination

An indirect enzyme-linked immunosorbent assay (ELISA) was developed to detect and differentiate the antibody responses to Japanese encephalitis (JE) virus nonstructural protein NS1 between infected and vaccinated individuals. The results showed that all convalescent sera from JE patients contained NS1-specific IgG antibodies, while 65 and 40% of these sera showed detectable NS1-specific IgM and IgA antibodies, respectively. Specificity analysis showed that NS1-specific IgM and IgA antibodies from JE patients do not cross-react to dengue virus NS1 glycoprotein, while IgG antibodies from 10% of JE patients showed significant cross-reaction to dengue virus NS1 glycoprotein. To differentiate infection from vaccination, the immune sera from 24 children vaccinated with inactivated JE vaccine were analyzed. The data showed that none of these immune sera had detectable NS1-specific IgG antibodies. The results demonstrated the potential application of JE NS1-specific indirect ELISA to differentiate infection from vaccination.     

Detection of antibodies to Japanese encephalitis virus in the wild boars in Hiroshima prefecture, Japan

Serum specimens were collected from 25 wild boars in Hiroshima prefecture located in the western region of Japan from November 2004 to February 2005. The sera were tested for antibodies to Japanese encephalitis virus (JEV) by IgM capture and IgG enzyme-linked immunosorbent assays (ELISA), and plaque reduction neutralization test. Seventeen samples (68%) were positive for neutralizing antibody to JEV. All the neutralizing antibody-positive samples were positive for IgG-ELISA. One was also positive for IgM. The results indicate that approximately 70% of the wild boars were positive for anti-JEV antibody, and raises the possibility that wild boars may play a role in the infectious cycle of JEV in this region.     

Comparison of serological methods for the detection of B. abortus antibodies in sera from vaccinated and non-vaccinated cattle.

A total of 4551 sera from 863 Strain 19 vaccinated and non-vaccinated adult cattle, independent of disease status, were tested by five serological methods to detect the presence of antibodies to B. abortus. Results from Standard Agglutination Tube (SAT), Buffered Brucella Antigen or card (CT), Complement Fixation (CF), Enzyme Linked Immunosorbent Assay (ELISA) and Rivanol (Riv) methods were compared. There was a 95% probability for agreement among CT negative sera, between serological methods, for all groups of vaccinated and non-vaccinated cattle. The agreement between tests with Riv Positive sera, excluding the calfhood and adult vaccinated group tested by the CF method, was 91-100%. The probability of a serum which was serologically negative by other methods being Riv negative was 98%. The usefulness of serological results from Riv (greater than or equal to 1/50) tests for classifying the reactor status of cattle are of doubtful supplemental value to confirm card test positive results. Vaccination history is an important consideration when evaluating serological data on cattle sera particularly from SAT and CF methods.     

江苏省自然人群乙型脑炎抗体血清学监测

目的了解江苏省正常人群流行性乙型脑炎(乙脑)免疫状况。方法采用蚀斑减少中和试验对2006-2009年收集的0～20岁血清标本3258份进行乙脑抗体检测。结果乙脑抗体总的阳性率为70.35%,GMT为1:13.56。结论江苏省3岁以上人群乙脑抗体有保护作用,其阳性率及效价随年龄增加而增加,0和1岁组抗体水平较低,应该加强该年龄段人群的保护。     

Roe deer as sentinels for endemicity of tick-borne encephalitis virus.

The suitability of serological surveys of roe deer (Capreolus capreolus) in determining the spread of tick-borne encephalitis virus (TBEV) was tested in a south German area with a low risk of TBEV infection to humans. Sera obtained from 192 hunted roe were screened by an haemagglutination-inhibition test (HAI) and in an ELISA developed in our laboratory. Those found positive were tested in a neutralization test (NT). Fifty (26.0%) sera reacted positive by ELISA and 43 (86.0%) of these were confirmed by HAI or NT. Forty-seven (24.5%) samples were positive by HAI, 44 (93.6%) of which were also positive in NT or ELISA. Only insignificant increase of the antibody prevalence with age (P = 0.17 for HAI antibodies) suggests that most infections occur at an early age in scattered natural foci. The antibody prevalence in females was lower than in males (OR = 0.63; P = 0.02 for HAI antibodies). In determining the distribution of seropositive roe we increased the sample size to 235 sera. No antibodies were detected in 56 (23.8%) sera collected in the eastern third of the county. The areas of high antibody prevalence in roe match those in which humans have been infected. We conclude that serosurveys of roe deer are useful in marking out areas in which humans face the risk of infection, provided that an adequate number of sera, preferably from males, is available.     

两种酶结合物诊断布鲁氏菌病的研究

以两种酶结合物(SPA-HRPO及兔抗人IgG-HRPO)做酶联免疫吸附测定(ELISA),对临床诊断为布鲁氏菌病人的血清51份进行了检査。结果除一例阴性外,其余50份均呈阳性反应。阳性效价为1:20~1:5120。用两种结合物检查结果基本一致,阳性率为98%。同时以凝集反应、2-ME试管凝集反应、补体结合反应虎红平板抗原凝集试验方法作了对比检查,阳性率分别为41%、35%、39%、31%。为了解ELISA法测定结果的特异性,使用混合布氏菌抗原吸收了被检血清,原来的阳性血清在吸收之后转成阴性。由于实验结果说明ELISA法作为布病诊断方法之一是可用的。