免疫酶染色试验检测斯氏狸殖吸虫抗体的研究

目的探索用免疫酶染色试验检测斯氏狸殖吸虫病抗体的敏感性和特异性。方法从病犬肺虫囊中收集斯氏狸殖吸虫成虫,冰冻切片,制作抗原片。采用免疫酶染色试验(IEST)检测斯氏狸殖吸虫病人和病鼠血清抗体。试验设健康大鼠血清对照及其他寄生虫病血清对照。结果斯氏狸殖吸虫病人和病鼠血清特异抗体阳性率分别为96.7%和97.4%。实验动物从感染后第2周开始抗体检测阳性,第4周阳性率达97.4%,持续10周。结论免疫酶染色试验敏感性高、特异性强,对斯氏狸殖吸虫病的早期诊断有重要参考价值。     

免疫酶染色试验检测斯氏狸殖吸虫抗体的研究

目的 探索用免疫酶染色试验检测斯氏狸殖吸虫病抗体的敏感性和特异性。方法从病犬肺虫囊中收集斯氏狸殖吸虫成虫，冰冻切片，制作抗原片。采用免疫酶染色试验（IEST）检测斯氏狸殖吸虫病人和病鼠血清抗体。试验没健康大鼠血清对照及其他寄生虫病血清对照。结果斯氏狸殖吸虫病人和病鼠血清特异抗体阳性率分别为96．7％和97．4％。实验动物从感染后第2周开始抗体检测阳性，第4周阳性率达97．4％，持续10周。结论免疫酶染色试验敏感性高、特异性强，对斯氏狸殖吸虫病的早期诊断有重要参考价值。     

流行性出血热患者唾液中特异性IgM抗体检测

本文报道应用IgM—抗体捕捉ELISA法检测流行性出血热患者唾液中特异性IgM抗体的结果。该法特异、敏感。检测出EHF患者发病第一天的唾液IgM抗体,阳性率92.86％(26/28)。其中21例在第15～20病日同时采集血清和唾液标本,检测结果唾液IgM阳性率80.95％(17/21)、将血清唾液IgM抗体均阳性的标本5例经2—ME试验,加热试验证明了其特异性。唾液标本较血清取材更方便、简单、成本低。     

用不同基因型戊型肝炎病毒重组蛋白建立酶联免疫吸附测定一步法

目的 以7种不同基因型和亚型的HEV重组蛋白作为包被抗原,建立HEV抗体检测ELISA一步法.方法 通过方阵滴定法确定包被抗原浓度,确定临界值,并进行敏感度、特异度和热稳定性试验.结果 7种重组抗原混合物(Mix166)最佳包被浓度为1.5 mg/L.抗-HEV IgG检测试剂的批内、批间变异系数分别为8.67%和10.85%,抗-HEV IgM检测试剂的批内、批间变异系数分别为4.56%和5.99%.一步法检测50份HEV RNA阳性血清的HEV IgG和HEV IgM抗体,阳性率均为94%.一步法检测674份健康者血清,52份抗-HEV IgG阳性,3份抗-HEV IgM阳性.一步法检测HEV墨西哥株攻击黑猩猩后收集的系列血清发现,病毒攻击后1～6周抗-HEVIgM阳性,2～76周抗-HEV IgG阳性,而进口试剂盒缺乏对抗-HEV墨西哥株IgG、IgM的反应性.结论 Mix166作为包被抗原建立的HEV抗体ELISA一步法具有较好的敏感性和特异性,可用于HEV感染的诊断.     

旋毛虫病患者血清中循环抗原及抗体检测的研究

111例旋毛虫病患者用斑点ELISA 试验检测循环抗原(CAg),薄板ELISA试验检测特异性抗体。其中未经治疗的患者36例,两法检出率均为100％;75例经治疗后3个月的患者血清中CAg 检出率为78.67％,抗体 IgG为 97.47％、IgM为 84.82％,IgE为 83.54％,两组间的 CAg、特异性 IgM、IgE抗体都有明显差异。健康人血清、日本血吸虫病患者血清试验,均有一定的假阳性与交叉反应。斑点ELISA检测旋毛虫病患者血清中CAg,ELISA检测IgM、IgE仍属敏感的和特异的,并且重现性好。特别是斑点ELISA采用第一和第二抗体标记酶进行两次放大的方法比之选用生物素亲和素放大法价廉,易于制备,从而可以取代传统的活检法,具有实用价值。     

抗体捕捉酶联免疫吸附法在流行性乙型脑炎早期诊断上的应用

用MacELISA测定了138份临床疑似乙脑病人的血清标本,并与HI试验进行了比较。MacELLSA能特异、敏感地检出乙脑患者发病第2天的IgM抗体,阳性率为55.56％。第4、5病日,IgM抗体阳性率高达92.86％。10例正常人和10份流行性出血热病毒抗体阳性的血清标本无一份阳性。     

班氏丝虫病人治疗前后血清抗体水平及免疫学诊断方法的研究

应用马来丝虫成虫冰冻切片抗原免疫酶染色技术(IEST),检测班氏丝虫微丝蚴阳性病人107例及治疗后一年微丝蚴阴转者35例和治后三年者112例血清特异IgG、IgE和IgM抗体水平,结果表明:治前病人血清IgG抗体滴度≥1:40者阳性率为96.3％,阳性GMRT为116.6;IgE抗体滴度≥1:20者阳性率为86.9％,阳性GMRT为37.1;IgM抗体滴度≥1:20者阳性率为88.8％,阳性GMRT为31.0,该三种抗体从阳性滴度     

弓形虫IgM-抗体捕捉酶联免疫吸附法的初步探讨

<正> 本文主要报道采用羊抗人IgM(n链特异)抗体包被,加受检血清,然后加弓形虫抗原和抗弓形虫特异性多抗的IgM-抗体捕捉酶联免疫吸附法(I_gM anti-body-capture ELISA)检测抗弓形虫特异性IgM的结果。该法检测肉类加工人员,献血员阳性率分别为44.7％×(38/85)和2.6％(1/36),而用常规间接IgM-ELISA阳性率分别为62.3％(53/85)和13.9％(5/36);同时与抗核抗体阳性血清和类风湿因子阳性血清的交叉反应阳性率,捕捉法为0％(0/36),间接法为0％和42.9％(15/36),并且对其中7份捕捉法检出IgM阳性的类风温因子阳性血清经2-ME吸收试验,结果确     

天津某医院严重急性呼吸综合征暴发期间血清流行病学调查

目的 研究本院严重SARS暴发流行后，疫源地不同人群血清SARS冠状病毒（SARS-CoV）抗体水平及其产生和变化规律；探讨人群SARS-CoV隐性感染和IgG抗体的保护作用。方法 采用血清流行病学法，联合运用ELISA和免疫荧光试验（IFA），调查疫情暴发流行期间及流行后，疫源地内非SARS人群血清抗体水平变化；定性研究SARS患者病后6周内IgM、IgG抗体产生和变化规律；动态观察SARS患者康复期82周内IgG抗体水平变化规律。结果 各100例疫源地一般人群和非疫源地对照人群血清SARS-CoV IgG抗体ELISA抽样检测均为阴性；487例SARS高危人群血清SARS-CoV IgG抗体ELISA检测阳性率为0．41％，经IFA复核后均为阴性；疫源地内非SARS人群IgG抗体A值水平在疫情流行后高于流行期间，差异有统计学意义（P〈0．05）；SARS患者病后1～6周血清抗体IgM和IgG的阳性检出率分别为7．7％、40．0％、57．8％、88．2％、76．6％、57．1％和0、23．1％、48．4％、65．4％、77．8％、100．0％；SARS患者康复期血清IgG抗体A值逐渐增高，病后第22周达到高峰，此后缓慢下降，第82周时仍维持较高水平，下降趋势减缓。结论 SARS可能不存在隐性感染者；绝大多数SARS患者在急性期或恢复早期的血清中存在IgM抗体，其出现较早，消失也较快，抗体IgG出现稍晚，但在血清中存在时间较长，达到高峰后消退缓慢，提示该抗体可能具有保护作用。     

日本血吸虫不同抗原检测家兔感染及治疗前后IgG/IgM抗体动态

目的探索特异性抗体检测在日本血吸虫病诊断和疗效考核中的潜在应用价值。方法ELISA方法采用可溶性虫卵抗原(SEA)、成虫抗原(AWA)检测人工感染家兔治疗前后血清中特异性IgGI、gM抗体,评价特异性抗体检测的诊断和疗效考核价值。结果用SEA抗原检测IgM抗体,发现感染后7周不管治疗与否,抗体水平均快速下降。与SEA相比,感染后AWA特异性IgM抗体上升时间早,且治疗后5个月仍为阳性。SEA特异性IgG抗体水平最高,但治疗后5个月仍为阳性。结论采用SEA抗原检测IgG用于血吸虫病诊断效果较好;对早期感染的诊断可以考虑SEA抗原检测IgM抗体作辅助。AWA抗原检测IgM,对急、慢性血吸虫感染均有较好诊断效果。采用这两种抗原检测IgG和IgM,均无考核疗效意义。     

联合检测血清抗日本血吸虫IgM和IgG4抗体意义的探讨

目的探讨联合检测血清特异性IgM和IgG4对日本血吸虫病的诊断价值。方法采用间接ELISA法检测急性血吸虫病、慢性血吸虫病及慢性血吸虫病患者治疗12月、24月后特异性IgM和IgG4的抗体变化。结果急性血吸虫病患者特异性IgM和IgG4阳性率分别是100%、87.9%,慢性血吸虫病患者特异性IgM和IgG4阳性率分别是45.5%、68.2%,联合检测急性血吸虫病和慢性血吸虫病患者特异性IgM和IgG4阳性率分别是100%和75.8%,慢性血吸虫病患者治疗12月、24月后特异性IgG4阳性率分别是16%、5%。结论并联检测特异性IgM和IgG4对急性血吸虫病早期诊断及慢性血吸虫病疗效考核具有较好的应用价值。     

IgM-ELISA法早期诊断广州管圆线虫感染的初步研究

目的探索广州管圆线虫病IgM早期诊断时机。方法从感染福寿螺中分离第Ⅲ期幼虫,感染6w龄Balb/c小白鼠,分别于感染前及感染后的第1、2、3、4、8、16、21、28、38d取血。经方阵滴定确定最佳抗原包被量和血清最佳包被浓度,进一步测定感染广州管圆线虫小鼠的不同时期血清中IgM含量,同时设立阴性、阳性对照。结果小鼠血清中IgM抗体在感染后就开始升高,感染后第8dIgM抗体检测阳性率已达100%。结论感染后1w,小鼠血清中IgM检测已具有临床诊断价值。     

胶体金免疫渗滤法快速诊断肾综合征出血热的临床研究

目的 建立一种快速，敏感的检测血清中抗汉坦病毒IgM抗体的方法。方法 将抗人IgM抗体点于硝酸纤维素膜上，以捕获患者血清中特异性IgM抗体，向已包被抗体的硝酸纤维素膜上滴加患者血清后，再选后滴加汉坦病毒抗原和鼠抗汉坦病毒抗体，最后滴加金标抗鼠抗体直接显色，阳性反应为红色，整个操作过程一般不超过10分钟。结果 用该法与IgM抗体捕获法ELISA对比检测51份肾综合征出血热（HFRS）患者血清标本，两法均阳性者49份，均阴性者2份。总符合率为100%，敏感性为96%。结论 胶体金免疫渗滤法操作简便，反应迅速，敏感性较高，适于在基层医疗单位中应用，可用于HFRS的早期诊断。     

Microtiter enzyme-linked immunosorbent assay for rubella antibodies in human sera--analysis by parallel line assay method]

Antibodies against rubella virus in human sera were measured by ELISA and antibody titers were calculated by the parallel line assay method. The dose response regression curves of standard sera and test sera containing IgG antibody calculated by the parallel line assay method showed linearity, and were parallel to one another. However, the regression lines of sera positive for IgM antibody against rubella virus were not parallel to one another in the low dilution region, but parallel in the higher dilution region. A good correlation was observed between the rubella IgG antibody titers measured by the hemagglutination-inhibition (HI) test and those calculated by the parallel line assay method. The coefficient of the correlation was 0.781. Time-course studies of IgG or IgM antibody titers against rubella virus in rubella patients or in vaccines with MMR vaccine indicated that the ELISA was more precise and specific method than the HI test. Thus, the parallel line assay method using ELISA is considered to be a more useful method for the detection and quantification of antibodies to rubella than the conventional HI test.     

ELISA test for antimeningococcal IgG and IgM antibodies: application to epidemiology and diagnosis.

We have developed an enzyme-linked immunosorbent assay (ELISA) in order to quantitate antimeningococcal IgM and IgG serum antibodies. The B:15 meningococcal strain was used as coating antigen, and class specific antibodies were detected by using alkaline phosphatase labelled rabbit anti-human IgM or IgG as conjugate. The specific IgG activity was higher in sera from healthy meningococcal carriers than non-carriers, but the difference was not statistically significant. Antimeningococcal IgM serum antibodies were more frequent in carriers that in non-carriers. Acute sera from 34 patients with fulminant meningococcal disease contained less specific IgG and had a higher prevalence of IgM than healthy carriers and non-carriers. By combining measurement of antimeningococcal IgG and IgM antibodies in both acute and convalescent sera 15/18 meningococcal patients demonstrated an increase in either IgG and IgM antibodies during the hospital stay, giving a sensitivity of 83%. 8/118 individuals without meningococcal disease had detectable specific IgM antibodies in their serum, giving a clinical specificity of the test of 93%. We conclude that quantitation of specific IgG antimeningococcal antibodies by a whole bacteria ELISA test may be a useful test for the study of immunity against meningococcal disease in single individuals as well as in epidemiological studies. The combined use of the IgG and IgM tests is helpful in the diagnosis of meningococcal disease when blood or cerebrospinal fluid cultures are negative.     

Rapid detection of IgM and IgG antibodies to herpesviridae virus

In this study, the immunoconcepts EA indirect enzyme antibody technique (colorzyme) was used not only for detection of IgG antibodies but also for quantitative detection of IgM antibodies to Herpes Simplex virus (HSV), Cytomegalovirus (CMV) and Epstein Barr Virus (EBV) to diagnose recent iactivei infection. Reference reactive and negative antisera and randomly collected human sera were tested by complement fixation test (CFT) against HSV antigens and tested also by immunofluorescent (IF) and colorzyme Immunoconcepts EA tests. All sera that were negative to HSV, CMV and EBV antibodies by CFT were negative by IF and colorzyme EA tests. All antibody positive sera and reference positive antisera were also positive by IF and colorzyme EA tests with slight variation in antibody titres between CFT and colorzyme test results. Human sera which were negative or IgM positive to HSV, CMV and EBV by ELISA as well as negative and positive reference sera from different diagnostic kits were retested by IF and colorzyme EA for IgM antiviral reactivity results were concordance by the three rests. All incubations in colorzyme test were at room temperature and only an ordinary microscope used in IF test or plate washers and readers needed for ELISA test. The colorzyme immunoconcepts is a simple, rapid and sensitive for viral diagnosis and can be used in any private laboratory.     

Immunoprecipitates used as a reagent for rheumatoid factors

IgM and IgG rheumatoid factors (RFs) were purified by affinity chromatography and gel filtration. The preparations thus obtained were used as standards in a radio-immunoassay for RF detection. In this assay, RFs were reacted with immunoprecipitates, and the RFs were detected with radiolabelled (F(ab')2 fragments specific for human IgM or IgG. The reproducibility of the assay was higher when RF content was expressed relative to the standards and not directly relative to tracer binding. It was found that the presence of IgM RF did not affect the measurement of IgG RF in this RIA, since the addition of neither mono- nor polyclonal IgM RF to a donor serum resulted in increased IgG RF measurements. 100 sera were analysed and were consistently positive in only one of the tests: sheep cell agglutination or latex fixation. The 75% of the sera which were positive only in the latex fixation test were positive for IgM RF in the radio-immunoassay, indicating that RFs in this type of serum were not specific solely for human IgG.     

Detection of IgM antibody against region IV flagellin of Salmonella paratyphi A

Salmonella paratyphi A is a pathogenic bacterium that causes paratyphoid fever. The current laboratory diagnostic techniques are unsatisfactory. To improve diagnosis, a plasmid (pSK-8E) encoding phase 1 flagellin gene nucleotide position 452-890 from S. paratyphi A has been constructed. The recombinant protein expressed from the plasmid has been used to develop an indirect ELISA for IgM antibody detection. Sera from patients with hemoculture positive for S. paratyphi A, S. typhi, other gram-positive and gram-negative bacteria, and dengue hemorrhagic fever as well as from healthy control subjects were tested. Sensitivity, specificity, positive and negative predictive values of the test were 56.9%, 98.8%, 90.6% and 92.1%, respectively. Since the sensitivity was low, the explanation for this result was investigated. It was found that the sensitivity of the test could be increased to 83.3% if the sera were obtained 9-12 days after onset of fever. The sera obtained earlier or later gave only 33.3% and 66.6% sensitivity, respectively. This result suggests that the IgM antibody detection assay which we have developed is a valuable tool for diagnosis of S. paratyphi A infection when the serum samples are taken at the appropriate time.