人抗狂犬病病毒IgG抗体3种检测系统的可比性研究

目的对比人抗狂犬病病毒IgG抗体定量测定酶联免疫试验(ELISA)试剂盒Autobio RABV-IgG ELISA、快速荧光灶抑制试验(RFFIT)以及BIO-BAD公司生产的PLATELIA~(TM)RABIESⅡKIT,检测狂犬病疫苗免疫后人血清抗狂犬病病毒IgG抗体水平的差异。方法收集狂犬病疫苗免疫前、免疫后临床血清样本共110份,使用RFFIT确定阳性标本(≥0.5 IU/ml)46份、阴性标本(0.5 IU/ml)64份,分别用Autobio RABV-IgG ELISA及PLATELIATM RABIESⅡKIT同时对人抗狂犬病病毒IgG抗体进行测定,对比3种检测系统对人抗狂犬病病毒IgG抗体检测结果的阴性符合率、阳性符合率及—致性。结果 PLATELIA~(TM)RABIESⅡKIT与RFFIT的符合率为92.7%;Autobio RABV-IgG ELISA与RFFIT的符合率为91.8%,与PLATELIA~(TM)RABIESⅡKIT的符合率为97.3%。结论 3种检测系统之间呈现良好的符合率,Autobio RABV-IgG ELISA可用于人抗狂犬病病毒IgG抗体检测。     

916例狂犬病疫苗免后血清抗体检测结果分析

916例暴露者接种狂犬病疫苗后15d采静脉血ELISA间接法检测人血清中抗狂犬病毒抗体,阳转率96.62%(885/916),阴性率3.38%(31/916),女阳转率分别为96.27%、97.05%,两组间无显著性差异(P>0.25),接种失败的31人再次全程免疫后28例阳转,3例仍为阴性。各年龄组间也无显著差异,建议暴露者接种疫苗后进行血清抗体检测,对抗体阴性者,应考虑重新全程免疫或加强免疫1~3针或给予免疫调节剂,以达到产生抗体的目的。     

蛋白印迹试验和第4代酶联免疫吸附试验艾滋病病毒检测结果

目的 对2011—2014年艾滋病病毒(HIV)抗体筛查阳性标本的确证试验结果进行分析。方法 2011—2014年收集天津市HIV高危人群的实验室资料,采用第4代酶联免疫吸附试验(ELISA)及胶体硒试验,对送检的和本筛查中心实验室检出的220例筛查阳性标本,使用蛋白印迹试验(WB)进行确证试验。结果 220例阳性标本,确证结果为阳性的病例180例(81.82%),经过1周,进行2次确证试验,阳性符合率为180例(100%)。阴性病例共10例(4.55%),不确定病例共30例(13.64%)。确证的180例中,gp160、gp120出现率达到100%,p55、p39阳性率为66%(119例)、61%(110例),gp41出现率为96%(173例),p51出现率为93%(167例),p31出现率为(160例),p24出现率为98%(176例)。结论 HIV抗体初筛试验结果存在假阳性,WB和第4代ELISA筛查试验可提高对艾滋病病毒的检查准确率,对WB试验中不确定标本需做好随访工作,避免艾滋病患者流失。     

连云港市2014－2016年健康人群流行性脑脊髓膜炎免疫水平和带菌率监测分析

目的 分析连云港市健康人群流行性脑脊髓膜炎免疫水平和带菌监测数据,为预防控制流行性脑脊髓膜炎提供参考依据。 方法 2014－2016年采集不同年龄组健康人群血标本和咽拭子标本,用流脑杀菌力实验检测血清流脑抗体,咽拭子进行流脑病原分离、玻片凝集实验。 结果 2014－2016年共采集不同年龄健康人群咽拭子标本1 265份,脑膜炎奈瑟菌阳性150份,带菌率11.86%。分群鉴定分离出B群2株,带菌率0.16%;C群3株,带菌率0.24%;其余群别未检出。带菌率最高的年龄组为3~岁,带菌率为18.95%,其次为15~岁(16.15%)。共采集1 239名健康人群血清标本,A群抗体阳性率76.84%,C群抗体阳性率82.97%。A群、C群3~岁及以上年龄人群抗体阳性率均高于80%。地区间、不同年龄组间A群、C群抗体阳性率和带菌率差异有统计学意义(P<或=0.001)。不同性别A群、C群抗体阳性率和带菌率差异无统计学意义(P>0.05)。地区间A群、C群抗体滴度差异有统计学意义(P<0.001)。 结论 连云港地区健康人群流脑A群、C群抗体阳性率较高。今后应加强B群流脑病例监测与防控,加强适龄儿童流脑A群、A+C群疫苗预防接种。     

IgG抗体亲和力实验在麻疹疑似病例检测中的应用研究

目的 使用 IgG抗体亲和力方法鉴别麻疹病例中的原发性和继发性免疫反应,研究麻疹消除阶段免疫失败病例的检出状况。 方法 选择出疹后3 d内和4~28 d采集的双份血清IgM结果均为阴性的麻疹疑似病例以及有明确免疫接种史的麻疹疑似病例,采用ELISA检测血清标本的IgG抗体,对于检出的IgG抗体进一步进行亲和力测定。 结果 52例双份血IgM阴性的麻疹疑似病例中,根据核酸或者IgG结果可确诊的麻疹病例有23例,其中19例IgG抗体为高亲和力,提示为再次感染。有明确含麻疹成分疫苗接种史的病例7例,其中1例病例接种一剂次含麻疹成分疫苗,采集的双份血中首份血产生低亲和力抗体,第二份血产生高亲和力抗体;其余6例接种≥2剂次含麻疹成分疫苗,均产生高亲和力抗体。 结论 IgG抗体亲和力检测可以发现再次感染的不典型麻疹病例,并能有效区分原发性和继发性免疫反应病例,为麻疹消除阶段的病例检测和控制提供技术支持。     

ELISA法检测放射工作人员风疹病毒感染的研究

目的 用一种快速检测抗风疹病毒抗体的诊断方法,了解放射工作人员风疹感染情况。方法 应用改良的间接ELISA法,对放射工作人员进行风疹病毒特异性抗体IgG和IgM检测。结果 济南地区514名放射工作人员中90.47%对风疹病毒具有抵抗力,6.42%为易感者,2.33%为原发感染,0.78%为再次感染风疹病毒。结论 放射工作人员的风疹抗体还存在一定的免疫空白,建议对他们开展疫苗免疫,消除风疹感染的危险性。     

反复呼吸道感染患儿食物特异性IgG抗体检测的临床意义

目的 探讨食物特异性IgG抗体检测在儿童反复呼吸道感染疾病中的临床使用意义。方法 选取开封市儿童医院2016年3月－2018年3月期间收治的门诊及住院经确诊的反复呼吸道感染患儿147例作为研究组,另选取同期进行体检的健康儿童110例作为对照组。采用酶联免疫法检测研究对象血清中牛肉、鸡肉、鱼、牛奶、蛋清/蛋黄、蟹、虾、大豆这8种食物特异性IgG抗体,分析两组儿童对食物特异性IgG的阳性率,并比较IgG阳性儿童血清中总IgE及嗜酸性粒细胞的阳性率。结果 研究组患儿的食物血清特异性IgG检出阳性率为73.5%,高于对照组正常儿童的18.2%(χ²=61.836,P<0.05);<3岁、3~<6岁及6~14岁年龄段,研究组的食物血清特异性IgG抗体阳性检出率分别为75.6%、81.3%及57.9%,均显著高于对照组(均P<0.05);研究组在牛奶、蛋清/蛋黄、蟹这三个食物过敏原的IgG检出阳性率为38.1%、57.8%、8.8%,均显著高于正常对照组(P<0.05);两组IgG阳性儿童中IgE及嗜酸性粒细胞检出阳性率结果差异无统计学意义(均P>0.05)。结论 食物特异性IgG抗体在反复呼吸道感染患儿中具有较高的阳性检出率,可以根据食物特异性IgG抗体的检测结果辅助反复呼吸道感染临床诊断和治疗。     

华容县1268例狂犬病疫苗接种后抗体水平分析

目的了解不同年龄、性别、季节、采血时间、免疫程序等因素对免疫效果的影响。方法采用间接ELISA法对2007年1月-2009年11月期间华容县疾病预防控制中心预防医学门诊采集到的检测狂犬病毒抗体的1268份血清标本进行检测。结果接种疫苗后,抗体的阳性率为93.7%,除季节、性别因素外,年龄、免疫程序采血时间均为血清抗体水平的影响因素。结论人被动物致伤后接种狂犬病疫苗,可获得有效的保护,免疫后检测血清抗体水平十分重要,对暂未产生抗体者要及时加强免疫提高抗体阳转率,严重咬伤者注射抗狂犬血清后,再进行全程免疫可获得满意的免疫效果,同时要注意采血时间对抗体检测结果也会产生影响。     

2017年北京市昌平区健康人群百日咳、白喉、破伤风抗体水平监测

目的 了解北京市昌平区健康人群百日咳、白喉、破伤风抗体水平,为制定免疫策略提供依据。方法 2017年采取多层抽样方法对360名健康人群采集血标本,采用酶联免疫吸附试验(ELISA)对血清进行百日咳、白喉、破伤风IgG抗体检测,分析不同年龄组、性别、户籍等之间抗体水平情况。结果 昌平区健康人群百日咳、白喉、破伤风抗体阳性率分别为5.00%、59.44%、68.33%,抗体几何平均滴度(geometric mean concentration,GMC)分别为9.18、0.55、0.86 IU/ml。不同年龄组白喉、破伤风抗体水平差异有统计学意义(P<0.001)。不同性别、户籍和免疫剂次人群间百日咳、白喉、破伤风抗体水平差异均无统计学意义(P>0.05)。接种末剂DTaP和/或DT和/或DTaP-IPV/Hib≤1年、2~5年、6~14年后的百日咳、白喉、破伤风抗体水平差异均有统计学意义(P<0.001)。结论 昌平区15岁以上人群白喉、破伤风抗体阳性率较低,随着免疫后时间延长,百日咳、白喉、破伤风抗体水平均明显降低。应加强小月龄婴儿、青少年及成人百日咳、白喉、破伤风的监测,推荐15岁以上人群、与1岁以下儿童有密切接触的成人及医务人员接种1剂Tdap疫苗。     

血清EB病毒抗体EA-IgA、VCA-IgA和Rta-IgG联合检测在鼻咽癌诊断中的意义

目的 探讨血清EB病毒EA-IgA抗体、VCA-IgA抗体和Rta-IgG抗体联合检测在鼻咽癌早期筛查和诊断中的临床价值。 方法 收集2019年初次就诊的264例鼻咽癌患者和264例健康体检者血清样本及其临床资料,采用酶联免疫吸附法(ELISA)检测血清中EA-IgA、VCA-IgA和Rta-IgG抗体水平,并评价各项指标在鼻咽癌诊断中的价值。 结果 EA-IgA、VCA-IgA和Rta-IgG抗体单独诊断鼻咽癌的灵敏度分别为57.20%、47.35%、75.38%;特异度分别为98.11%、97.35%、96.21%。其中Rta-IgG抗体的灵敏度最高,特异性较好。初诊鼻咽癌组三种抗体的血清学水平或阳性率均显著高于健康对照组,差异均有统计学意义(P<0.05)。在联合检测诊断鼻咽癌时,EA-IgA+VCA-IgA+Rta-IgG抗体三项联合的灵敏度最高,为92.05%;AUC面积最大,为0.946;阴性预测值最高,为92.02%;漏检率最低,为7.95%;特异度好,为91.67%。EA-IgA、VCA-IgA和Rta-IgG抗体阳性率与性别、年龄、临床分期和TNM分期均无明显相关性(P>0.05)。 结论 EA-IgA+VCA-IgA+Rta-IgG抗体三项联合检测对于鼻咽癌早期筛查及诊断具有更高的临床价值。     

狂犬病暴露后使用“2-1-1"程序接种人用狂犬病疫苗(Vero细胞)3年免疫持久性及2剂加强免疫效果观察

  目的  评价使用"2-1-1"程序接种狂犬病疫苗的免疫持久性及2剂加强免疫效果。  方法  选择曾暴露后使用"2-1-1"程序后1年、2年和3年的对象314人,进行2剂加强免疫,在加强免疫前、后14天分别采集血清,采用酶联免疫吸附实验(enzyme linked immunosorbent assay,ELISA)法检测人狂犬病毒IgG抗体,分析使用"2-1-1"程序后1年、2年、3年及经2剂加强免疫后的抗体几何平均浓度(geometric mean concentration,GMC)和抗体阳性率。  结果  303人按"2-1-1"程序接种疫苗后1年、2年和3年的抗体GMC分别为1.33 IU/mL、1.04 IU/mL和0.72 IU/mL,抗体阳性率分别为77.78%、66.67%和55.56%。282人经2剂加强免疫后,1年组、2年组和3年组的抗体GMC分别为16.83 IU/mL、19.37 IU/mL和21.05 IU/mL,加强免疫后抗体GMC均高于加强免疫前(t=16.54,P < 0.001;t=13.85,P < 0.001;t=16.02,P < 0.001);抗体阳性率分别为100.00%、99.00%和100.00%。  结论  狂犬病暴露后使用"2-1-1"程序具有良好的免疫持久性,3年内2剂次加强免疫后具有较好的免疫应答。     

53例人用狂犬病无佐剂纯化疫苗接种后抗体检测结果分析

目的观察暴露后人群接种国产人用狂犬病无佐剂纯化疫苗(Vero cell)的免疫效果。方法对53例研究对象按照0、3、7、14和28 d程序进行暴露后免疫,采用WHO认可的RFFIT法检测免疫14 d,45 d的血清抗体。观察每针次接种后72 h内局部和全身反应。结果所有接种对象均未出现严重副反应。首剂免疫14 d,45 d ELISA法检测抗体阳性率分别为52.83%,94.34%,RFFIT法检测抗体阳性率分别为100%,100%。几何平均滴度(GMT)为17.74 IU/ml,15.50 IU/ml。结论国产人用狂犬病无佐剂纯化疫苗(Vero cell)具有良好的安全性和免疫原性。     

Standardisation and establishment of a rabies ELISA test in European laboratories for assessing the efficacy of oral fox vaccination campaigns

A simple and rapid enzyme linked immunosorbent assay (ELISA) to detect rabies antibodies in field fox sera has been standardised and established in several European laboratories. The same panels of 100 coded sera were investigated by four laboratories using this ELISA assay and reference serum neutralisation techniques (fluorescent antibody virus neutralisation (FAVN) test and rapid fluorescent focus inhibition test (RFFIT)). This indirect ELISA technique is highly correlated with conventional seroneutralisation test on cell culture. Results obtained with this ELISA test provide an almost perfect agreement between laboratories while the agreement of FAVN test and RFFIT results is substantial and very satisfactory. In view of this study, this simple and rapid ELISA test is a suitable tool for evaluating the sero-conversion rate in fox populations following oral rabies vaccination campaigns in European countries.     

Evaluation of a new serological technique for detecting rabies virus antibodies following vaccination

Two major techniques are currently used to estimate rabies virus antibody values: neutralization assays, such as the rapid fluorescent focus inhibition test (RFFIT), and enzyme-linked immunosorbent assays (ELISAs). The RFFIT is considered the gold standard assay and has been used to assess the titer of rabies virus neutralizing antibodies for more than three decades. In the late 1970s, ELISA began to be used to estimate the level of rabies virus antibody and has recently been used by some laboratories as an alternate screening test for animal sera. Although the ELISA appears simpler, safer and more efficient, the assay is less sensitive in detecting low values of rabies virus neutralizing antibodies than neutralization tests. This study was designed to evaluate a new ELISA-based method for detecting rabies virus binding antibody. This new technique uses electro-chemi-luminescence labels and carbon electrode plates to detect binding events. In this comparative study, the RFFIT and the new ELISA-based technique were used to evaluate the level of rabies virus antibodies in human and animal serum samples. By using a conservative approximation of 0.15 IU/ml as a cutoff point, the new ELISA-based technique demonstrated a sensitivity of 100% and a specificity of 95% for human samples and for experimental animal samples. The sensitivity and specificity for field animal samples was 96% and 95%, respectively. The preliminary results from this study appear promising and demonstrate a higher sensitivity than traditional ELISA methods.     

间接免疫荧光技术检测狂犬病抗体IgG

目的：研究间接免疫荧光技术在检测狂犬病抗体IgG中的应用。方法：研制狂犬病抗原片和生产荧光血清，研究间接免疫荧光技术检测狂犬病抗体IgG的灵敏度、稳定性及实际应用效果。用研制的狂犬抗原片和荧光血清，比较间接免疫荧光技术与酶标检测狂犬病抗体的效果。结果：间接免疫荧光技术检测狂犬病抗体IgG是特异、稳定的，变异系数CV=4．9％。间接免疫荧光技术与ELISA同时检测狂犬病抗体，间接免疫荧光法阳性率为92．89％，ELISA法阳性率为40．48％。结论：间接免疫荧光法测定狂犬病抗体IgG特异性、稳定性好。狂犬疫苗免疫后应用该技术检测抗体，保证免疫效果。     

Evaluation of an improved rapid neutralizing antibody detection test (RAPINA) for qualitative and semiquantitative detection of rabies neutralizing antibody in humans and dogs

Using the principle of immunochromatography, we previously developed a method called RAPINA (Rapid Neutralizing Antibody detection test) that can measure the level of rabies virus -neutralizing antibody (VNA) in serum samples [Shiota S, Mannen K, Matsumoto T, Yamada K, Yasui T, Takayama K, et al. Development and evaluation of a rapid neutralizing antibody test for rabies. J Virol Methods 2009;161:58-62]. RAPINA is faster, simpler, and easier to perform compared with a virus-neutralizing test or enzyme-linked immunosorbent assay (ELISA). The improved version of RAPINA has greater positive and negative predictive values corresponding to a VNA level of 0.5 IU/mL, as recommended by the World Health Organization and the World Organization for Animal Health. To verify the efficacy of this improved method, serum samples were collected from humans and dogs before and after immunization against rabies and were tested in Japan, Sri Lanka, and Thailand. The results were compared between RAPINA and the true VNA levels measured by the Rapid Fluorescent Focus Inhibition Test (RFFIT). The improved RAPINA accurately predicted seropositivity for 182 of 183 seropositive human samples as assessed by RFFIT (99.5%) and for 138 of 140 RFFIT-negative human samples (98.6%). In dog serum samples, the positive and negative predictive values were 99.7% (345/355) and 95.6% (174/182), respectively. RAPINA was also used to estimate VNA levels in a semiquantitative manner by using serial dilution of serum samples. Our results show that RAPINA is an easy and rapid method for measuring VNA levels before and after immunization with the rabies vaccine and does not need a high skill level or sophisticated equipment. RAPINA can be used to monitor the success of preexposure prophylaxis in at-risk persons, vaccine coverage, and animal control. It can also be used in laboratories with modest facilities and where a large number of samples are screened.     

北京市891例狂犬病疫苗接种后抗体水平分析

目的:了解疫苗种类、免疫程序、采血时间、年龄、性别等因素对免疫效果的影响。方法:采用间接ELISA法对2007年北京市疾病预防控制中心免疫门诊采集到的检测狂犬病毒抗体的891份血清标本进行检测。结果:接种疫苗后抗体阳性率为92.7%,除性别因素外,年龄、接种疫苗的种类、免疫程序、采血时间均为血清抗体水平的影响因素。结论:绝大多数人群在接种狂犬病疫苗后可获得有效的保护,抗体阳转率在一定范围内随接种针次的增加而提高,严重咬伤者注射抗狂犬病血清后再进行全程免疫可获得满意的免疫效果。同时要注意采血时间对抗体检测结果也会产生影响。     

Multicenter comparative study of a new ELISA, PLATELIA RABIES II, for the detection and titration of anti-rabies glycoprotein antibodies and comparison with the rapid fluorescent focus inhibition test (RFFIT) on human samples from vaccinated and non-vaccinated people

The envelope glycoprotein G of rabies virus induces the production of neutralising antibodies, which are important in protection against rabies. Therefore, titration of anti-envelope glycoprotein antibodies is a good indicator of the degree of immunity in people during anti-rabies treatment or after vaccination. According to the World Health Organization (WHO) guidelines, a booster vaccine dose should be given if the rabies antibody titre falls below 0.5 IU/ml. Titration of anti-rabies antibodies is also useful for plasma centers in the preparation and standardization of human anti-rabies gamma-globulins for therapeutic use and to a lesser extent for the diagnosis of rabies in human sera and cerebrospinal fluid (CSF). This paper presents a new enzyme-linked immunosorbent assay (ELISA), PLATELIA RABIES II, developed for rabies envelope glycoprotein antibody detection or titration and its comparison to the current reference method (RFFIT). The data collected during validation of the test in a multicenter study are analysed to give a sound overall knowledge of the capabilities of the PLATELIA RABIES II, for instance specificity, linearity, accuracy, precision, detection limit and quantitation limit. To this aim, human serum samples from a total of 1348 vaccinated or non-vaccinated people were tested in parallel using the new ELISA and the RFFIT for the presence of anti-rabies antibodies. Data generated indicate a linear relationship across the range of titration between the two methods. The sensitivity reaches 98.6% and the specificity 99.4%. This study indicates that this new ELISA test is as sensitive and specific as the current standardized reference method. The method is simple, safe, rapid and can be considered as a useful alternative to the neutralisation test.     

Comparative evaluation of specific ELISA and RFFIT antibody assays in the assessment of dog immunity against rabies

Two techniques are currently used to evaluate the humoral immune responses to rabies vaccination: ELISA, which detects binding antibodies to viral antigens and the WHO reference rapid fluorescent focus inhibition test (RFFIT), which assays in vitro virus-neutralizing antibodies. In this study, we have comparatively evaluated antibody responses of dogs reared either in an experimental kennel or living in field conditions after vaccination with a cell culture-derived rabies vaccine. In experimental conditions, both ELISA and RFFIT techniques were well correlated. However, in field conditions, they yielded discrepant results particularly in evaluating the residual rabies immunity before vaccine administration and in identifying seroconverted dogs. After rabies vaccination in field conditions, while similar antibody titres and seroconversion rates were obtained using either technique, the discrimination of a given dog according to the seroconversion threshold depended on the assay. We concluded, that whereas in experimental conditions, ELISA and RFFIT were well correlated, in field conditions ELISA yielded upper estimates. Consequently, RFFIT, although a cumbersome test, should continue to be considered as the reference rabies antibody assay technique. A seroconversion threshold of 0.5 IU/ml should be cautiously considered and a higher threshold (1 IU/ml) could be more appropriate in the evaluation of rabies immunity in the field in order to marginalize the interfering factors.     

2013-2016年天津市狂犬病暴露后免疫效果分析

目的 了解天津市狂犬病暴露后免疫者血清中和抗体水平,为指导狂犬病预防控制工作提供科学依据。方法 对2013-2016年在天津市疾病预防控制中心动物致伤门诊全程注射狂犬病疫苗并于免疫后14～30天采血进行快速荧光灶抑制试验者进行统计学分析。结果 不同月份接种狂犬病疫苗,中和抗体水平趋于平稳,在10IU/ml上下波动;未成年组头面部暴露比例最高,青壮年组下肢暴露比例最高,老年组上肢暴露比例最高;91例暴露后免疫者血清中和抗体几何平均滴度为9714IU/ml,阳转率为100%;女性中和抗体水平明显高于男性(t=2482,P=0015);复种组中和抗体水平远高于初种组(疫苗组及疫苗+蛋白组)(F=5356,P=0006);不同年龄组、不同暴露部位、不同暴露等级及接种不同疫苗中和抗体水平无统计学差异(P皆＞005)。结论 狂犬病暴露后免疫效果不受季节、年龄、暴露部位、暴露等级及接种疫苗种类影响,女性免疫效果优于男性,复种组免疫效果明显优于初种组。