SARS病毒抗体的检测及其应用

为了解SARS患者血清抗体产生的规律 ,我们用间接免疫荧光法 (IFA)和酶联免疫吸附试验 (ELISA)对广州市第八人民医院的 4 3例住院治疗的SARS患者和 10例正常人的双份血清进行SARS抗体检测 ,同时比较了IFA和ELISA法的特点。10例正常人双份血清的SARS病毒IgM和IgG抗体均阴性。 4 3例SARS患者 ,其中 34例检出IgM占 79.1%。 37例检出IgG占 86 .10 %。SARS病毒IgG抗体的阳性率高于IgM。结果显示 ,发病的同时机体便可检测到SARS抗体IgM最迟离起病时间最长 (33d)的血清也能检测到 ,但也有患者发病第 9、2 3及 30天的第二份…     

麻疹IgM抗体行定量酶联免疫吸附试验检测应用效果分析

目的探讨对麻疹患者采用定量酶联免疫吸附试验测定IgM抗体的临床价值。方法回顾性分析在我院接种麻疹疫苗的100例儿童的临床资料,分别采用定量酶联免疫吸附试验和中和抗体试验测定血清IgM抗体水平,比较两者的符合率;同时统计ELISA与NT诊断的耗时、费用和阳性率。结果浓度≥200 IU/L样品ELISA检测与NT的符合率明显高于200 IU/L样品,其差异有统计学意义(P0.05)。两种检查手段阳性率无明显差异(P0.05),但ELISA法的耗时和费用明显低于NT法其差异有统计学意义(P0.05)。结论定量酶联免疫吸附试验在检测麻疹Ig M抗体时具有很高的准确性,且耗时和费用均较低,值得临床推广。     

20例SARS患者特异性抗体变化规律

目的：了解严重急性呼吸道综合征(SARS)特异性IgM和IgG抗体的变化规律。方法：采用间接酶联免疫吸附试验(ELISA)检测20例SARS患者系列血清中特异性IgM和IgG抗体，系列血清包括患者发病后1周，2周，3周，4周，8周，12周所采集的样本。结果：20例患者发病后第1周IgM和IgM抗体均为阴性；第2周时16例IgM抗体阳性，17例IgM抗体阳性；第3周后所有患者IgG抗体阳性并持续至第12周，而IgM抗体阳性患者逐渐减少，至第12周时所有患者均为阴性。结论：SARS特异性IgG抗体消失较早，其存在是近期感染的标志；IgG抗体的持续存在可能是获得病后免疫力的标志。     

应用MAC-ELISA方法检测呼吸道合胞病毒IgM抗体

建立酶联免疫吸附试验(MAC-ELISA)检测呼吸道合胞(RS)病毒特异性IgM抗体方法,并与中和试验(NT)进行对比。 取35例患毛细支气管炎婴幼儿的急性期及恢复期双份血清共70份标本进行研究。MAC-ELISA测得急性期血清中RS病毒特异性IgM抗体的阳性率为27/35(77.14％),而恢复期血清中和抗体呈≥4倍升高者为23/35(65.71％)。凡NT阳性病例,MAC-ELISA也呈阳性;凡MAC-ELISA阴性病例,NT也呈阴性。另4例NT阴性而MAC-ELISA呈阳性。两者阳性符合率为88.57％。 本研究也观察了IgM抗体与年龄的关系。在6个月龄以前,MAC-ELISA的阳性率远远高于NT。 实验说明采用MAC-ELISA检测RS病毒特异性IgM是可靠的快速诊断方法。     

SARS患者康复期血清特异性抗体效价检测

目的：SARS患者康复期血清特异性抗体的效价。方法：采用间接酶联免疫法和双抗原夹心法。结果：SARS患者康复期血清特异性抗体IgG水平在住院第5周达最高值，IgM水平在住院第3周达到最高值。SARS患者康复期血清IgG水平是IgM的1～7倍。住院5～7周的SARS康复期患者，其血清特异性抗体检测阳性率达100％。结论：SAPS患者康复期血清特异性抗体水平在住院第5周达到最高值，其阳性检出率达100％。     

Ⅰ型单纯疱疹病毒特异性IgG、IgM及中和抗体的消长动态

本文采用酶联免疫吸附试验(ELISA)间接法、固相抗 IgM ELISA及微量中和试验对 HSV-1感染家兔特异性 IgG、IgM 及中和抗体的消长动态进行观察。结果表明,特异性IgM 在初次感染后第4天可检出,第8～15天达峰值,第66天完全消失,再感染后则检测不出。特异性 IgG 及中和抗体在初次感染后第 8～15天出现,第22天达峰值,检出时间长达 186天以上,再感染后滴度骤升50～100倍。     

武汉地区128例乙型脑炎患者特异性IgM抗体的检测及临床分析

本文应用IgM抗体捕获酶联免疫吸附试验(MAC ELISA)对武汉地区1986～1987年128例临床诊断符合流行性乙型脑炎(简称乙脑)患者的血清及脑脊液进行JEV IgM检测,并对检测结果及阳性、阴性病例的临床特点进行了分析、讨论,现报告如下。     

甘肃省SARS抗体血清流行病学调查

目的 了解甘肃省严重急性呼吸综合征(SARS)患者、密切接触者和正常人群血清SARS冠状病毒特异性IgG抗体水平。方法 利用酶联免疫吸附试验(ELISA)法检测SARS病毒IgG抗体水平。检测对象包括甘肃省9例SARS患者的急性期和(或)恢复期系列血清，l109例直接护理SARS患者的医生、护士、实验室检测人员、疾控人员和曾与患者有接触的人员以及978例正常人血清。结果 9例临床诊断病例SARS冠状病毒特异性IgG抗体中6例为阳性，疾病恢复后12个月血清抗体仍为阳性；密切接触者1例特异性IgG抗体阳性；正常人3例特异性IgG抗体阳性。结论 病例抗体阳性符合临床诊断，密切接触者和正常人的抗体阳性数较低，提示可能不存在隐性感染。     

类风湿关节炎患者CMV、B19和HCV抗体的检测及临床意义

目的:检测巨细胞病毒(CMV)IgG和IgM抗体、细小病毒B19 IgG和IgM抗体及丙型肝炎病毒(HCV)抗体在中国南方类风湿关节炎(RA)患者外周血中的表达,探讨CMV、B19、HCV在RA中的作用。方法:采用酶联免疫吸附试验(ELISA)法检测CMV-IgG和CMV-IgM抗体,B19-IgG和B19-IgM抗体以及HCV抗体;CMV-IgM抗体阳性者利用实时定量PCR法检测外周血单个核细胞中的CMV载量;B19-IgM抗体阳性者利用实时定量PCR法检测血清中的B19载量。同时分析这些病毒抗体与RA患者的实验室活动指标如抗环瓜氨酸肽(CCP)抗体、类风湿因子(RF)、红细胞沉降率(ESR)的相关性。结果:70例RA患者中,69例为CMV-IgG抗体阳性(98.57%),7例CMV-IgM抗体阳性(10.00%),49例B19-IgG抗体阳性(70.00%),16例B19-IgM抗体阳性(22.86%),无一例HCV抗体阳性;92例健康对照者中91例CMV-IgG抗体阳性(98.91%),1例CMV-IgM抗体阳性(1.10%),42例B19-IgG抗体阳性(45.65%),19例B19-IgM抗体阳性(20.65%),仅1例HCV抗体阳性;RA患者CMV-IgM抗体阳性率明显高于对照组(P<0.05),CMV-IgM抗体阳性RA患者外周血单个核细胞未检测到CMV载量;RA患者B19-IgG抗体阳性率明显高于对照组(P<0.01),而两组间B19-IgM抗体阳性率无差别(P>0.05),B19-IgM抗体阳性RA患者血清中未检测到B19载量;CMV-IgM抗体阳性、B19-IgG抗体阳性与抗CCP抗体、RF、ESR等RA活动性指标不相关。结论:中国南方人群普遍感染CMV,但CMV重新激活与RA有关;B19感染在RA患者中有更高的流行率;HCV在中国南方人群感染率低。     

不孕及反复流产患者血清抗心磷脂抗体的检测

目的检测不孕及反复流产患者血清中的抗心磷脂抗体(ACA),探讨ACA的临床意义.方法应用酶联免疫吸附(ELISA)法检测105例原发或继发不孕患者(不孕组)、171例反复流产或有胚胎停育史患者(流产组)及40例正常对照组血清中的IgG、IgM及IgA-ACA 3种抗体.结果不孕组ACA总阳性率为44.76%,流产组为35.09%,均显著高于对照组(P＜0,01).不孕组及流产组的单纯IgG阳性率亦明显高于对照组(P＜0.05);而两组的单纯IgM阳性率与对照组相比,差异均无显著性意义(P＞0.05).结论ACA是导致不孕及反复流产的免疫学因素之一,IgG型的ACA比IgM型的更具临床意义     

流行性出血热特异性循环免疫复合物的动态观察和分析

应用双抗体夹心ＥＬＩＳＡ检测了８８例流行性出血热患者２２８份血清中特异性循环免疫复合物，并与胶固素－ＥＬＩＳＡ检测循环免疫复合物的结果进行了比较。结果表明，胶固素－ＥＬＩＳＡ检出循环免疫复合物１８３份（８０．２６％），抗原特异性ＥＬＩＳＡ检出ＩｇＧ型循环免疫复合物１８８份（８２．４５％），ＩｇＭ循环免疫复合物２０５份（８９．９１％），ＩｇＡ型循环免疫复合物１４７份（６４．４７％）。在各类特异性循环免疫复合物中，ＩｇＭ型在各病期检出率最高，ＩｇＧ型次之，ＩｇＡ型最低。在发病初期，各类特异性循环免疫复合物检出率均较高，极期达高峰，随病情缓解而渐降低。对特异性循环免疫复合物检出率与流行性出血热患者病情程度相关性的观察表明，轻型患者检出率较中，重型患者为低，而中、重型间检出率差别不明显。此外，轻型患者特异性循环免疫复合物动态变化明显，极期达高峰后很快下降，而中、重型患者检出率下降趋势较缓慢。上述结果直接证实了流行性出血热循环免疫复合物的形成是特异性病毒抗原刺激机体免疫应答的结果，并构成了流行性出血热免疫发病机制中的重要环节。     

Serological evidence of dengue fever among refugees, Hargeysa, Somalia

Epidemics of a malaria-like illness affected several thousand residents of the Dam Camp, a refugee camp near Hargeysa in Somalia, during 1985, 1986, and 1987. The disease was characterized by fever, chills, sweats, headache, back and joint pains for as long as 10 days in some patients. Blood smears from acutely ill patients were negative for malaria. Of 28 acute and 10 convalescent sera tested by the indirect fluorescent antibody (IFA) and by the hemagglutination inhibition (HI) tests, all were negative for antibody to Rift Valley fever, Crimean-Congo hemorrhagic fever, Sindbis, Chikungunya, yellow fever, and Zika viruses. However, antibody reactive to dengue 2 virus was detected by the IFA test in 39% (15/38), and 11 of 29 (38%) of the same sera were antibody positive by the HI test. Also, IgG antibody reactive to dengue 2 was demonstrated in 60% (17/28) of the same sera by the enzyme immunoassay (EIA), and 14% (4/28) were positive for IgM antibody. Of ten patients for which acute and convalescent sera were available, two developed four fold or greater rises in antibody titer evidencing infection. These data suggested that dengue virus may have been the cause of the epidemic among the Dam Camp refugees.     

流行性出血热循环免疫复合物各组分的动态观察及意义

应用ＰＥＧ沉淀法对２７例１１４份流行性出血热病人血清中循环免疫复合物进行了检测，不同病期循环免疫复合物检出率分别为：发热期８８．８％，少尿期８２．５％，多尿期４７．８％，恢复期２１．４％。对流行性出血热循环免疫复合物组分的动态观察发现，在不同病期循环免疫复合物中绝大多数可检出流行性出血热病毒抗原及免疫球蛋白与补体Ｃ３，表明在流行性出血热患者体内检出的循环免疫复合物是特异性病毒抗原刺激机体免疫应答的     

用免疫转印技术检测流行性出血热病人特异性抗体的探讨

采用免疫转印技术检测了147份临床诊断为EHF病人血清中的特异性IgG抗体,结果133份血清可检出针对分子量在68和66kd左右两种病毒多肽的抗体,检出率为90.4％,略高于免疫荧光试验(88.4％),两种抗特异性多肽抗体的几何平均滴度分别为荧光法的4倍和2.1倍。本法具有特异性强、敏感性高和重复性较好等特点,并能检出针对不同抗原亚基成份的抗体,可作为EHF特异性诊断的方法之一。     

Hemadsorption immunosorbent technique for determination of mumps immunoglobulin M antibody

We used hemadsorption immunosorbent technique (HIT) to detect mumps immunoglobulin M (IgM) antibody. IgM from human sera was adsorbed into anti-human IgM-coated wells in plates, and mumps-specific IgM was detected by adding mumps virus hemagglutinin and guinea pig erythrocytes consecutively. Specific IgM-positive sera showed hemadsorption, whereas negative sera showed hemagglutination. All 81 patients with current mumps infections tested showed mumps-specific IgM antibody, with titers ranging from 160 to 327,680. In most cases IgM antibody was already present on day 1 or 2 after the onset of illness. IgM antibody persisted for 6 to 10 weeks. Of 57 patients with acute respiratory illnesses caused by parainfluenza virus, 5 showed cross-reactions in the hemadsorption immunosorbent technique assay for mumps IgM. The hemadsorption immunosorbent technique assay is specific for the IgM class of antibody and avoids false-positive results due to rheumatoid factor. This test is an efficient and sensitive method for rapid and early diagnosis of mumps infections.     

An amplified ELISA for the detection of parvovirus B19 IgM using monoclonal antibody to FITC

A new M-antibody capture ELISA for the detection of specific IgM against parvovirus B19 is described. The test uses a monoclonal anti-fluorescein isothiocyanate (FITC) antibody conjugated to horseradish peroxidase to amplify the positive reactions. Serum samples (N = 823) submitted for B19 IgM assay were tested in parallel in the new ELISA and the standard B19 M-antibody capture radioimmunoassay (MACRIA) test. By both tests B19 IgM was detected in 38 (4.6%) samples, and not detected in 771 (94%) samples. One sample was positive in the ELISA test but negative in the RIA and of the 13 sera giving 'equivocal' results in the MACRIA, 6 were positive and 7 negative. If the RIA equivocal results were excluded, the ELISA showed 100% (38/38) sensitivity and 99.9% (771/772) specificity compared to the MACRIA test. The B19 IgM ELISA is a sensitive and specific test with better discrimination between anti B19 IgM positive and negative specimens than MACRIA.     

Diagnosis of parainfluenza virus infection in children and older patients by detection of specific IgM antibody

The significance of detecting specific antibody of the IgM class for the diagnosis of parainfluenza infections was examined. Paired sera from 763 children and adults admitted to the hospital for acute respiratory disease were tested for significant antibody titer rises in the hemagglutination inhibition (HI) test and for specific IgM antibody with the hemadsorption immunosorbent techniques (HIT). Sera were collected during two 6-month periods, January through June, 1982 and 1983. Evidence of parainfluenza infections was found in 122 patients (16%): 83 (25%) in 1982 and 39 (9.1%) in 1983. The HIT was superior to the HI test for detection of parainfluenza infection, in particular in infants and aged patients, 94 patients were positive only in the HIT test, 12 in the HI test, and 16 in both tests. In a control group of 120 persons (time- and age-matched to the patients of 1982) admitted for nonrespiratory illness, six (5%) showed parainfluenza IgM in their serum. Blocking experiments and retrospective clinical information indicated that the IgM antibody detected in these individuals is specific IgM acquired after a mild parainfluenza infection. Most (66%) patients showed IgM antibody titer rises or high titers (greater than 1,280) in both sera, and in 23%, a fall in IgM antibody titer was found. Detection of specific IgM antibody by HIT permitted early presumptive diagnosis in 71% of the patients with parainfluenza infection. IgM antibody persisted for 2-11 weeks. The HIT appears to be an important supplement for the diagnosis of parainfluenza infections.     

Laboratory diagnosis of primary and secondary dengue infection.

BACKGROUND: Dengue fever is routinely detected in many laboratories using commercial tests for the specific detection of dengue IgM antibodies. OBJECTIVES: We have studied the sensitivity of IgM antibody detection in paired serum samples of 43 patients with either with primary dengue (PD) or secondary dengue (SD). STUDY DESIGN: Two consecutive samples were drawn from 23 Vietnamese and 20 German patients. All patients were selected for a positive PCR and for the fact that consecutive serum samples were available. The diagnosis of PD was based on seroconversion to dengue antigen and in SD on the detection of virus RNA in the presence of anti-dengue IgG antibodies. RESULTS: In samples of patients with PD fever taken during days 1-3 of the disease no IgM antibody could be detected. During days 4-7 and after day 7, IgM antibody was detected in 55% and 94%, respectively. In patients with SD fever, even less positive IgM samples were found in samples taken during days 4-7 (47%) and after day 7 (78%). IgG titers were significantly higher in SD compared to PD patients, although high (>1280) titers were also found in some PD patients. CONCLUSION: In numerous acute dengue fever patients an early diagnosis will be obtained only by combining IgM antibody detection with detection of virus or virus RNA using RT-PCR.     

Persistence of specific IgM and low avidity specific IgG1 following primary rubella.

Persistence of specific IgM in sera following primary rubella infection was compared with the maturation of the specific IgG1 response. 206 sera, from 171 patients with primary rubella, taken 1 day to 2.5 years after onset of illness, were tested. Rubella-specific IgM was detected by M-antibody capture radioimmunoassay in 100% of sera taken 15-28 days after onset, but in only 9% taken 3-4 months after onset. However, using the diethylamine (DEA) shift value (DSV) method, low avidity specific IgG1 was detected in 91% sera taken at 3-4 months and at 5-7 months 21% of sera remained positive. Using an avidity index method, with urea in the wash buffer, none of the sera were positive for low avidity specific IgG1 beyond 3 months after onset. With DEA in the wash buffer, the number of sera positive rose to 38% at 3-4 months. Thus, the DSV method for detecting low avidity specific IgG1 is a useful additional test for confirming or refuting a diagnosis of primary rubella and is of particular value for assessing pregnant patients.     

固相酶联免疫测定法检测NS1抗原在登革病毒感染早期诊断中的应用

目的探讨固相酶联免疫测定（ELISA）法检测NS1抗原在登革病毒感染早期诊断中的应用价值。方法选取登革病毒感染早期患者血清171份,非登革病毒感染发热患者血清11份,正常人血清10份,采用ELISA法检测全部192份血清的登革病毒NS1抗原和IgM抗体;采用逆转录-聚合酶链反应-限制性内切酶酶切片段长度多态性分析（RT-PCR-RFLP）技术对发病5 d内的125份血清进行扩增和鉴定分型;并采用C6/36细胞微量培养法对发病第1、2天的41份血清进行登革病毒分离培养。结果登革病毒感染患者发病2 d内、3～5 d以及6～10 d血清NS1抗原的检出率分别是92.7%（38/41）、83.3%（70/84）、10.9%（5/46）;IgM抗体的检出率分别是2.4%（1/41）、51.2%（43/84）、97.8%（45/46）;非登革病毒感染的发热患者及正常人血清中,有1例疟疾患者血清登革病毒IgM抗体呈阳性,NS1抗原无一例阳性。RT-PCR在登革病毒感染患者发病第1、2天和3～5天的检出率分别是85.4%（35/41）、83.3%（70/84）;登革病毒感染患者发病第1、2天血清的病毒分离培养阳性率分别是80.0%（16/20）、38.1%（8/21）,总分离率58.5%（24/41）;RT-PCR-RFLP分型鉴定技术及间接免疫荧光法（IFA）均证实2006年广州流行株为登革Ⅰ型病毒。结论ELISA法检测登革病毒NS1抗原操作技术成熟,且具有敏感性高、特异性好的特点,对登革病毒感染的早期诊断和疫情的早期控制具有重要意义,适合于基层医疗机构常规应用。