诊断丙型肝炎病毒感染的3种ELISA的比较研究

将新建立的检测血清中抗丙型肝炎病毒（ＨＣＶ）总抗体的双抗原夹心ＥＬＩＳＡ（夹心法）与普遍采用的检测抗ＨＣＶ－ＩｇＧ的间接ＥＬＩＳＡ（ＩｇＧ间接法）及检测抗ＨＣＶ－ＩｇＭ的间接ＥＬＩＳＡ（ＩｇＭ间接法）进行比较研究。检测１５份ＩｇＭ间接法阳性、ＩｇＧ间接法阴性（ＩｇＭ单阳）血清及２０份ＩｇＧ间接法阳性、ＩｇＭ间接法阴性（ＩｇＧ单阳）血清,夹心法均为阳性,这３５份血清经ＰＣＲ检测ＨＣＶＲＮＡ均阳性。夹心法可同时检出ＩｇＧ和ＩｇＭ抗体,检出率高于后两者。检测２０份ＨＣＶＲＮＡ阴性血清,３种方法均阴性。用生物制品检定所检测ＨＣＶ－ＩｇＧ抗体的质控血清为标准品,夹心法和ＩｇＧ间接法均可达到检测标准     

伯氏疏螺旋体基因工程抗原ELISA法检测莱姆病特异IgG …

目的：利用伯氏疏螺旋体基因工程抗原Ｐ３９建立间接ＥＬＩＳＡ法检测莱姆病特异性抗体ＩｇＧ。方法：基因工程抗原Ｐ３９的包被浓度和酶二抗所用浓度及血清稀释倍数,均由棋盘试验确定。并进行了精密度,特异性试验。阻断试验。免疫复合物溶解试验。干扰试验。结果：基因工程抗原Ｐ３９最佳浓度为２００ｎｇ．ｍｌ＾－１,用ＥＬＩＳＡ法鉴定２,Ｐ３９蛋白与两株Ｐ３９单克隆抗体均发生特异性反应。测定９０例血清,其中５０例阳性     

360例孕妇及其新生儿巨细胞病毒感染的血清学调查分析

本文采用酶免疫吸附试验（ＥＬＩＳＡ）法，检测３６０例孕妇及其新生儿血清中的巨细胞病毒特异性抗体ＩｇＧ和ＩｇＭ。孕妇血清ＣＭＶＩｇＧ。ＩｇＭ抗全的阳性率分别为６１．１％和３．１％。新生儿脐血ＣＭＶＩｇＧ和ＩｇＭ抗体的阳性率分别为５８．３％和１．４％。ＩｇＭ抗体阳性母亲所生新生儿体重低于ＩｇＭ阴性母亲所生儿的体重。５例新生儿ＩｇＭ阳性者中，１例脑积水畸形，１例先天性心脏病。证明孕妇ＣＭＶ感染可引起新生     

重组日本吸虫成虫32kDa抗原用于疗效考核的研究

目的探讨重组日本血吸虫成虫３２ＫＤ抗原（ｒｓｊ３２）检测日本血吸虫病人的特异性和敏感性及考核疗效的价值。方法采用ｒＳｊ３２－ＥＬＩＳＡ法检测治前及治后不同时期血吸虫病人血清中特异性ＩｇＧ及正常人、肺吸虫、肝吸虫和囊虫病人血清中交叉抗体。结果ｒＳｊ３２在敏感性和特异性方面与ＳＥＡ比较差异无显著性（Ｐ＞０．０５）;用ｒＳｊ３２－ＥＬＩＳＡ检测治后１２个月血清ＩｇＧ的阴性率为７６．７％,显著高于ＳＥＡ－ＥＬＩＳＡ的４３．３％,检测治后２４个月血清ＩｇＧ的阴性率达９０．５％。结论ｒＳｊ３２－ＥＬＩＳＡ检测日本血吸虫病具有与ＳＥＡ－ＥＬＩＳＡ相同的诊断价值和较高的考核疗效价值。     

用金标免疫斑点法检测抗精子抗体

抗精子抗体（ＡｓＡｂ）检测对免疫性不育的诊断具有重要意义 ,研究表明 ,在多种导致不育因素中 ,以抗精子抗体最为密切。抗精子抗体是由精子或精子膜抗原诱发的特异性抗体 ,它具有凝集精子细胞 ,抑制精子通过宫颈粘液向宫腔内移动 ,抑制精子获能、顶体反应及受精 ,从而降低生育力。其在血清中的存在通常以ＩｇＧ、ＩｇＭ为主 ,精浆中通常以ＩｇＡ、ＩｇＧ为主。目前应用的ＡｓＡｂ检测方法有 :精子制动试验（ＳＩＴ）、精子凝集试验（ＳＡＴ）、混合凝集试验（ＭＡＴ）、酶联免疫吸附试验（ＥＬＩＳＡ）、间接免疫荧光试验（ＩＩＦ）等 ,由…     

汉坦病毒核蛋白基因表达及产物抗原活性的研究

为了制备肾综合征出血热（ ＨＦＲＳ） 诊断用基因工程抗原,构建了合汉坦病毒７６１１８ 株编码核蛋白（ ＮＰ） 基因片段的重组质粒ｐＢＶ２２０Ｈ１－３Ｓ, 经转化大肠杆菌和诱导表达,ＳＤＳＰＡＧＥ 显示菌体中有分子量约为４８ ＫＤ 的新蛋白质生成。用经包含体纯化获得的表达产物做抗原,以间接酶联免疫（ＥＬＩＳＡ） 检测了４８ 份疑似出血热患者的血清特异性ＩｇＧ,其结果与经典的间接免疫荧光（ＩＦＡ） 检测的阴性、阳性及总符合率分别为１００ ％ 、９２－８６ ％ 和９３－７５ ％ 。证实该重组ＮＰ 具有良好的生物活性和诊断抗原价值     

斑点免疫金渗滤试验检测血清中抗登革病毒IgM抗体的研究

斑点免疫金渗滤试验检测血清中抗登革病毒ＩｇＭ抗体的研究姚海军段馥琴曹虹广州市卫生防疫站病毒免疫室（５１００８０广州）抗登革病毒（ＤＥＮＶ）ＩｇＭ抗体的测定常用ＭａｃＥＬＩＳＡ，但是，该法目前国内还没有试剂盒供应，国内使用者一般靠进口的ＭａｃＥＬＩＳ...     

肾综合征出血热ELISA诊断抗原的制备

为研制肾综合征出血热（ＨＦＲＳ）ＩｇＭ ＥＬＩＳＡ诊断试剂盒，本文改进了ＨＦＲＳ抗原的制备方法，即将感染出血热病毒（ＨＦＲＳＶ）的全细胞破碎添加以硫柳汞、牛血清、ＥＤＴＡ等配制的防腐保护剂，制备ＥＬＩＳＡ液相抗原。经不同温度、不同时间热稳定性试验，结果表明：抗原加保护剂经３７℃１个月，４℃９个月，－１５℃１２个月存放后，仍满足ＥＬＩＳＡ诊断抗原滴度要求。     

用ELISA检测大鼠血清中肾综合征出血热病毒特异性抗体

本研究应用单克隆抗体（ＭｃＡｂ）技术，建立了三种检测大鼠血清中肾综合征出血热病毒（ＨＦＲＳＶ）特异性抗体的ＥＬＩＳＡ。通过与间接免疫荧光法（ＩＦＡＴ）对比试验发现，三种ＥＬＩＳＡ在检出大鼠血清ＨＰＲＳＶ特异性抗体的阳性率和敏感性上均明显高于ＩＦＡＴ试验，并且ＥＬＩＳＡ与ＩＦＡＴ总检出符合率为８８．５％。此外还发现由于引入小鼠抗大鼠Ｋａｐｐａ轻链ＭｃＡｂ，抗体捕捉ＥＬＩＳＡ和间接夹心ＥＬＩＳＡ具有早期诊断的特性，这些方法的建立对野生及实验动物的ＨＦＲＳＶ流行病学监测及检疫有一定价值。     

不同年龄组痘苗病毒IgG抗体水平的调查研究

用ＥＬＩＳＡ方法检测了１６５例不同年龄组正常人血清中痘苗抗体水平，结果１２岁以下儿童痘苗病毒ＩｇＧ抗体全部阴性，１３岁以上各年龄组痘苗ＩｇＧ抗体阳性率在５３．８～８０．０％左右，阳性抗体效价分布在１∶２００～１∶１６００之间，几何平均滴度为１∶２６９～１∶２９５之间。虽然３１岁～４０岁年龄组痘苗ＩｇＧ抗体阳性率略有升高，但各年龄组之间痘苗抗体几何平均滴度无显著差异（Ｐ＞０．０５）。用间接免疫荧光方法检测，结果１～１２岁年龄组痘苗病毒ＩｇＧ抗体全部阴性，与ＥＬＩＳＡ方法检测结果一致，１３岁以上１００例血清中痘苗抗体阳性数２０例，阳性率２０％，但不同年龄组之间阳性率有所变化，但无明显差异。     

Antibody-capture enzyme-linked immunosorbent assays that use enzyme-labelled antigen for detection of virus-specific immunoglobulin M, A and G in patients with varicella or herpes zoster.

Antibody-capture enzyme-linked immunosorbent assays (AC-ELISA) which use enzyme-labelled antigen were developed for detection of varicella-zoster virus-(VZV) specific IgM, IgA and IgG antibody in patients with varicella or herpes zoster and in sera from healthy individuals. All 18 patients with varicella developed a VZV-IgM and a VZV-IgG response, 17 also a VZV-IgA response. In contrast, all 19 patients with herpes zoster were shown to be positive for VZV-IgA whereas only 13 of these reacted positively for VZV-IgM. A VZV-IgM response was detected in only two sera from 100 healthy individuals and an IgA response in only one. The presence of virus-specific IgA and IgG in the cerebrospinal fluid as determined by AC-ELISA was a useful indicator of VZV infection of the central nervous system. By AC-ELISA, VZV-IgG was detected predominantly in sera from patients with acute or recent VZV infection. Only 14 sera from 100 healthy individuals were positive for VZV-IgG by AC-ELISA, whereas all were positive by an indirect ELISA. These results indicate that AC-ELISA's may be useful assays for determination for acute or recurrent VZV infection, but are not suitable for determination of past infection with this virus.     

检测血清幽门螺杆菌细胞空泡毒素抗体的间接ELISA法初步建立

目的:以重组细胞空泡毒素(VacA)蛋白为抗原初步建立检测血清VacA抗体间接ELISA法,为制备检测Hp毒株感染的试剂盒奠定基础.方法:Ni-NTA2 树脂纯化重组VacA蛋白,用重组蛋白免疫兔,Helico Blot试剂盒检测兔血清VacA抗体以鉴定其免疫原性.Western blot检测其抗原性.并以纯化的重组蛋白为抗原,建立检测血清VacA抗体间接ELISA法,以Helico blot试剂盒为标准,确定该方法的特异性与敏感性.结果:以重组蛋白为抗原建立的检测血清VacA抗体的间接ELISA法,敏感性与特异性分别是93.1%和92.5%.结论:以重组蛋白为抗原初步建立检测血清VacA抗体的间接ELISA法,敏感性、特异性高,为制备商品化的试剂盒奠定了基础.     

巨细胞病毒特异性IgG抗体检测方法的建立及应用

目的 为了建立一种快速、简便、实用的可用于育龄妇女血清中巨细胞病毒(CMV)-IgG抗体的筛查方法,并初步探索其应用价值. 方法 建立重组CMV(pp150+52)抗原;以国家人口和计划生育委员会科学技术研究所(简称本所)建立的重组CMV(pp150+52)抗原作为包被抗原,700 IU/L作为临界值,用间接ELISA方法,对深圳宝安区1 024例血清标本进行CMV-IgG抗体的检测. 结果 该方法 灵敏度为13 IU/L,批内变异系数4.7%～7.0%,批间变异系数7.6%～14.4%,并与SORIN公司生产的试剂盒进行了比较,2种方法 呈最度相关,Kappa值为0.82;深圳宝安区1 024例血清标本CMV-IgG抗体阳性率为93.3%(955/1 024),阴性率为6.7%(69/1 024). 结论 本所建立的ELISA方法 快速、简便和实用,可用于育龄妇女CMV-IgG抗体的筛查.     

酶联免疫吸附试验间接抗体夹心检测SARS-CoV抗原方法的建立和应用

目的制备和鉴定严重急性呼吸综合征冠状病毒(SARS-CoV)核衣壳(N)蛋白单克隆抗体(rnAb)和多克隆抗体建立SARS—CoVN抗原捕获抗体夹心酶联免疫吸附试验(ELISA)方法用于SARS-CoV感染的早期诊断。方法用基因重组SARS-CoV N蛋白免疫BALB/c小鼠和新西兰大白兔制备mAb和多克隆抗体，采用ELISA间接法、免疫荧光和免疫印迹进行筛选和鉴定，用单克隆抗体与兔多克隆抗体进行配对试验，建立抗原捕获抗体夹心ELISA法测定SARS-CoV N抗原。结果获得9株特异性针对SARS-CoV N蛋白的mAb和高效价的兔多克隆抗体，通过高亲和力的mAb与兔多抗的配对试验，筛选出3株单抗N1E8、N8E1和N10E4混合作为捕获抗体，与兔多克隆抗体和辣根过氧化物酶标记羊抗兔IgG组合作为测定抗体，建立了抗体夹心ELISA法，测定重组SARS-CoV N蛋白最高灵敏度为50pg／ml，特异性达99．86％，测定420份血清学确诊的SARS患者血清，其中发病1—10天阳性检出率为90.1％，11—20天检出率为23％，21天以上均为阴性，与其他呼吸道病毒和冠状病毒无交叉反应。结论获得特异性好、亲和力高的单克隆抗体和高效价的兔多克隆抗体，经过抗体的配对和优化，建立了一种灵敏度高、特异性强的SARS-CoV抗原的ELISA捕捉法．可应用于SARS早期诊断、溯源及流行病学研究。     

应用12D5和21B7单克隆抗体检测广州管圆线虫循环抗原的研究

目的 研制快速诊断广州管圆线虫感染的金标层析检测试剂.方法 双抗体(12D5和21B7)夹心ELISA检测实验感染广州管圆线虫的大鼠、广州管圆线虫感染病例血清循环抗原(CAg),同时将12D5和21B7单抗点样于固相的硝酸纤维膜上,以胶体金-protein A为标记物制备金标免疫层析试剂快速检测实验感染广州管圆线虫的大鼠、广州管圆线虫感染病例血清循环抗原.结果经鉴定单抗12D5为IgG1,21B7为IgM,两株单抗同时识别广州管圆线虫成虫相对分子质量55×10~3的蛋白,12D5和21B7双抗体夹心ELISA和金标层析法对实验感染广州管圆线虫的大鼠血清中CAg检出率为100%(48/48),广州管圆线虫感染病例血清CAg检出率为100%(32/32),与日本血吸虫、肝吸虫、肺吸虫、旋毛虫、蛔虫、包虫病例血清无交叉反应,与健康人血清无反应.结论 12D5、21B7双抗体夹心ELISA和金标层析法对感染广州管圆线虫人和动物血清中CAg检测的特异性强,金标层析法操作简便、快速,结果判读容易,不需特殊设备,且敏感性高,能够确定现症感染.     

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.     

巴楚县2001年新疆出血热疫情的血清学证实

目的：以血清流行病学方法调查新闻出血热（XHF）病人，易感人群和主要宿主动物中疾病的感染情况。方法：分别收集2001年4－6月新疆巴县临床诊断为XHF的病人血清，易感人群血清和主要宿主动物的血清，用研制的诊断试剂以酶联免疫吸附试验（ELISA）检测XHF特异性IgG和IgM抗体；用抗原捕获ELISA检测XHF病毒抗原。结果：病人血清IgG抗体阳性率为39．62％（21／53）。IgM抗体阳性率为20．75％（11／53），抗原获ELISA有1份血清为XHF抗原阳性；易感人群血清IgG抗体阳性主为21．05％（4／19），IgM抗体检测和抗原捕获ELISA全部为阴性；羊血清IgG抗体阳性率为70％（56／80）。结论：血清流行病学研究证实该次疫情确系XHF、流行地区人畜均有较高水平的隐性感染。     

Leptospirosis serodiagnosis by ELISA based on recombinant outer membrane protein

The outer membrane protein LipL21, LipL32, LipL41 and Loa22 of Leptospira interrogans serovar Copenhageni were previously revealed by immunoproteomic analysis, using sera from acute phase infection in a guinea pig. The full-length DNA of each protein was then cloned from the same serovar and expressed in pRSET vector. The obtained molecular weight (MW) of recombinant proteins rLipL21, rLipL32 and rLoa22 were slightly higher than the MW predicted from nucleotide sequences of each inserted gene, while only the N-terminal half of rLipL41 was obtained. Mice antiserum raised against each purified recombinant protein could react with the whole cell lysate of leptospiral serovars, implying that leptospiral native proteins shared a common epitope with recombinant protein. Serodiagnosis using recombinant protein antigen based on indirect ELISA procedure was developed in this study. The optimization of the ELISA components lead to determination of optical density (OD) from a single serum-dilution of 1:1000 in the leptospirosis patients group and normal healthy control group. The cut off OD values for both IgG and IgM class were investigated, and based on this fixed dilution only the IgG class could be used for differential diagnosis of patients and normal individuals. Compared with the MAT assay, ELISA assay utilizing both rLipL32 and rLoa22 as antigen, gave high accuracy and could thus be useful as a confirmative serology test.     

Enzyme linked protein-A: an ELISA for detection of amoebic antibody

Enzyme linked protein-A was used to develop an enzyme linked immunosorbent assay (ELISA) system for the detection of circulating antibodies to amoebic antigen. The specificity of protein-A to bind IgG only through Fc receptors, makes the test more specific for the detection of IgG antibodies to amoebic antigen. The ELISA system was used to detect amoebic antibody in control subjects (56), patients with amoebic liver abscess (79) and Entamoeba histolytica cyst-passers (10) and the results compared with those of indirect haemagglutination assay (IHA). The ELISA was more sensitive and detected 74.7% of cases with amoebic antibody in amoebic liver abscess compared with 66.7% detected by IHA. The test was more specific, sensitive and easy to perform and is recommended as a test of choice for the serological diagnosis of amoebic liver abscess.     

Infections due to the human herpesviruses in southern India: a seroepidemiological survey

We studied the prevalence of IgG and IgM antibodies to the human herpesviruses in a hospital-based population of 181 individuals aged 0 to 25 years, who were resident in Vellore, south India or surrounding rural areas. Antibodies to the Epstein-Barr virus (EBV) viral capsid antigen were determined by indirect immunofluorescence, while antibodies to the remaining herpesviruses were determined by enzyme-linked immunosorbent assay. The age-specific prevalence rates of IgG antibodies to EBV and cytomegalovirus (CMV) rose rapidly after birth to reach a value of over 90% by the fourth year of life. High age-specific IgM prevalence rates and geometric mean titres (GMT) of IgG antibody in children 6 months to 2 years of age, and the early median age of virus infection (1.4 years for EBV and less than 1 year for CMV) indicate that primary infection with these viruses occurs early in life. In contrast, age-specific prevalence rates of IgG antibodies to varicella-zoster virus (VZV) and herpes simplex virus (HSV) rose gradually after birth to attain maximal values of only 72% (VZV) and 83% (HSV) in the 15-25 year age group, and the median ages of infection were delayed (12.25 years for VZV and 8.2 years for HSV). The age-specific IgG prevalence rates of VZV and HSV, and of EBV and HSV showed statistically significant positive correlations, suggesting that common epidemiological factors may underlie the pattern of infections due to these groups of viruses.(ABSTRACT TRUNCATED AT 250 WORDS)