日本血吸虫门脉内童虫表膜抗原组分及其保护性免疫力研究

目的探讨日本血吸虫门脉内童虫表膜抗原(SjHmAg)的免疫特性,观察其抗日本血吸虫(Sj)的保护效果。方法用SDS-PAGE电泳技术分析SjHmAg蛋白组分,酶联免疫电转移印迹(EITB)分析感染兔血清(IRS)和正常兔血清(NRS)对SjHmAg的识别;用完整SjHmAg免疫昆明鼠3次,分别在0、2、4周进行,第6周每鼠经腹部感染40±1条Sj尾蚴,42天后剖杀,计数虫数及肝卵数。结果用SDS-PAGE电泳获得SjHmAg主带7条,IRS主要能识别SjHmAg23、33和63kDa等10个抗原组分;间接ELISA测其抗体滴度>1:6400,与对照组相比,SjHmAg免疫小鼠的减虫率为16.2％,减卵率为55.4％。结论用SDS-PAGE获得了不同分子量的SjHmAg蛋白,EITB鉴别出具有免疫活性的蛋白分子,且SjHmAg对Sj攻击感染及雌虫生植似有一定的抗性。     

酶联免疫印迹术（EITB）在小鼠日本血吸虫感染早期诊断中的应用

为探讨日本血吸虫感染的早期诊断方法,本文应用酶联免疫印迹术(EITB)检测感染鼠血清的循环抗原。所用抗体由日本血吸虫成虫和虫卵的五种抗原(SEA、UEA、TA、CY、MC)免疫动物获得。SDS—PAGE和EITB显示这些抗原与华支睾吸虫、肺吸虫和蛔虫等交叉反应不明显。其中CY抗原的31KD条带是血吸虫特有。用EITB监测小鼠感染日本血吸虫后6周内各期和用吡喹酮治疗后2周的循环抗原有如下变化:UEA抗体显示的187KD抗原多肽在4周出现并逐渐加深,治疗后变淡;与TA和CY抗血清反应的40KD抗原分别在感染后3天～4周和3天～2周出现;抗MC显示的15.3KD在6周出现,治疗后消失;与成虫表膜相关的20、15、12.8KD仅在治疗后出现。对这些特异的抗原性多肽动态观察似可为早期诊断日本血吸虫病和考核疗效提供参考。     

实验日本血吸虫病早期循环抗原及抗体的动态

为了解实验动物感染日本血吸虫后早期循环抗原及抗体的发展动态,本文以竞争抑制型酶联免疫吸附试验(I-ELISA)观察循环抗原的变化及酶联免疫及附试验(ELISA)观察抗体变化。 I-ELISA所用5种抗原,包括成虫3种——皮层抗原(TA)、胞浆抗原(CY)及微粒体抗原(MC),虫卵2种——可溶性抗原(SEA)及尿素溶性抗原(UEA).     

用Sephadex G200层析血清对4种弓形虫病免疫诊断方法的评价

弓形虫感染小鼠血清经Sephadex G 200柱层析后,各层析峰(紫外线OD_(280)吸收峰)用染色试验(DT)、乳胶凝集试验(LAT)、间接荧光抗体试验(IFA)和酶联免疫吸附试验(ELISA)分别检测总抗体、IgM和IgG抗体。结果显示,两类抗体分别存在于第Ⅰ峰和第Ⅱ峰,第Ⅲ峰无抗体活性。ELISA和IFA用于弓形虫病早期诊断(检测IgM)具有高度特异性和敏感性;LAT操作简便、经济快速、但敏感性稍差。IFA可取代DT用于常规个例诊断。     

日本血吸虫各期虫体的酶联免疫电转移印迹分析及抗原定位研究

本文采用酶联免疫电转移印迹技术（EITB）分析、比较日本血吸虫不同发育时期虫体特定的组分蛋白分子、雌虫、雄虫和虫卵抗原分别与相应的雌、雄虫和虫卵免疫血清反应呈现17、20和8条蛋白带,这三种抗原与其它不同时期虫体免疫血清反应,三者间及彼此间均出现交叉反应,而雌、雄虫和虫卵抗原与其它寄生虫感染血清作用没发现有叉及反应。应用间接荧光抗体试验（IFA）和免疫酶染色试验（IES）对日本血吸虫抗原进行定位研究,其结果相似。血吸虫主要抗原物质来源于成虫表皮、肠上皮和卵内的毛蚴.     

人脐带血中丝虫抗原的IgM抗体:经胎盘感染的可能性

本文报告了用间接免疫荧光试验(IFA)、酶联免疫吸附试验(ELISA)和固相放射免疫试验(SRIA)对母体和脐带血中抗丝虫抗体的检测结果。     

IFA和ELISA检测正常人及SLE患者血清SARS-CoV抗体特异性比较

为探讨酶联免疫吸附试验(ELISA)和间接免疫荧光试验(IFA)检测严重急性呼吸综合征冠状病毒(SARS—CoV)抗体的特异性，2003年5～8月，我们对此进行了有关研究。现报告如下。     

对一种快速筛选刚地弓形体抗体免疫测定法的评价

检测特异性抗刚地弓形体(Toxoplasma gondii)抗体的价值很大,已研制出许多抗体检测方法。这些方法如Sabin-Feldman染色试验、间接血凝试验(IHA)、间接免疫荧光试验(IFA)、酶联免疫吸附试验(ELISA)及荧光免疫测定在大型临床实验室中容易使用,但不适于快速筛选。作者评价了一种市售的一次使用诊断系统(Single Use Diagnostio System;SUDS)检测T.gondii抗体的性     

肿瘤患者血清学弓形虫抗体检测方法的比较

<正> 弓形虫病最可靠的诊断方法是确认病原,但一般认为比较困难。因此,应用血清学方法检测弓形虫抗体已被公认是重要的辅助诊断手段。我们选择了常用的间接血凝试验(IHA)、间接荧光抗体试验(IFA)以及斑点酶联免疫吸附试验(Dot-ELISA),对202例肿瘤患者做了弓形虫抗体检测,并进行了初步的比较研究。 被检血清取自天津肿瘤医院。IHA所用弓形虫抗原购自兰州兽医研究所,批号691205。操作步骤按IHA常规方法进行,血清滴度>1:64判为阳性。IFA抗原用RH株弓形虫接种小鼠72小时后,取腹水,经处理,涂片固定凉干,密闭储存于4℃备用。荧光素标     

两种试剂盒快速检测斯氏狸殖吸虫抗体的研究

目的观察肺吸虫病金标渗滤试剂盒(DIGFA kit)和酶联免疫吸附试验(ELISA)试剂盒检测斯氏狸殖吸虫抗体的敏感性和特异性。方法采用DIGFA kit和ELISA检测斯氏狸殖吸虫病流行区人群和病鼠血清抗体。结果 DIGFA和ELISA试剂盒检测斯氏狸殖吸虫病流行区人群和病鼠血清特异性抗体阳性率均为5.08%(54/1062)和100%(40/40);用两种试剂盒检测正常大鼠、旋毛虫病大鼠、血吸虫病兔和蛔虫病病人血清,除1例血吸虫病兔血清DIGFA出现阳性反应外,其他血清两种方法均为阴性。大鼠感染斯氏狸殖吸虫后第2周用两种方法检测抗体,阳性率分别为20.00%(8/40)和27.50%(11/40),第4周阳性率均达100%,并持续至第8周。结论 DIGFA试剂盒具有与ELISA试剂盒相同的敏感性和特异性,且有操作快速、简便、直观等特点,适合在基层和斯氏狸殖吸虫病流行地区推广使用。     

Evaluation of enzyme linked immunosorbent assay in the immunodiagnosis of schistosomiasis.

Eighty-six bilharzial patients divided into 5 clinical groups were studied. Enzyme linked immunosorbent assay (ELISA), indirect fluorescent antibody (IFA) and indirect haemagglutination (IHA) test were performed for all the patients. ELISA gave the most sensitive results (82.6% and 80.2% positivity rate in Egypt and Lille respectively), followed by IFA (79.1% positivity rate) and IHA (77.9% and 75.6% positivity rate in Egypt and Lille respectively). The humoral antibodies detected by all methods (ELISA, IFA and IHA) showed increasing values with the progress of the disease which is parallel to the antigenicity of the disease. Both the positivity rate and the mean value of antibody titres recorded by the 3 diagnostic techniques (ELISA, IFA and IHA) were significantly higher in mansoni than haematobium infection. This may be explained by species specificity of antibody response. The superiority of ELISA over IFA and IHA techniques was discussed.     

Epidemiological survey of Trypanosoma cruzi infection in north-eastern Brazil using different diagnostic methods.

A survey of the prevalence of Trypanosoma cruzi infection was carried out in Oitis, a small community in the State of Piaui, Brazil. Two hundred and sixty five individuals were screened by microscopic examination, hemoculture, indirect immunofluorescence (IFA), enzyme-linked immunosorbent assay (ELISA), and competitive enzyme-linked immunosorbent assay (C-ELISA) using the monoclonal antibody TCF87 against to a 25kd T. cruzi antigen. Seropositivity was 14.3% by the IFA test, 14.7% by ELISA, and 13.2% by C-ELISA. The C-ELISA using the TCF87 monoclonal antibody seems to be applicable in serodiagnosis of Chagas' disease.     

Serodiagnosis of canine visceral leishmaniasis in Portugal: comparison of three methods.

Sera collected in Portugal from 43 dogs were screened for specific antibodies to Leishmania donovani antigens. Three different techniques were compared: an indirect immunofluorescence assay (IFA), a direct enzyme-linked immunosorbent assay (ELISA) and a competitive-ELISA (C-ELISA) using two species-specific monoclonal antibodies, D2 and D13. By IFA, 22 of the sera examined showed positive reactions, compared with 26 by ELISA or 27 by C-ELISA. There was no direct correlation observed between the serum titre by IFA and the strength of the reaction in ELISA or inhibition in C-ELISA. However, a good correlation was observed between sera identified as positive (95.5%) by all three techniques. Western blotting on leishmanial membranes showed that common antigens with Mr of 26,000 and 70-84,000 were recognized by all infected dog sera, regardless of the serum titre. In large scale studies, ELISAs are preferred to IFA for the rapid diagnosis of canine visceral leishmaniasis because of their greater simplicity.     

Prevalence and comparison of serologic assays, necropsy, and tongue examination for the diagnosis of porcine cysticercosis in Peru

Swine cysticercosis, a severe zoonotic disease which is part of the Taenia solium life cycle, causes major economic losses in pig husbandry. Throughout South America, farmers diagnose cysticercosis by examining the tongues of their pigs for cysticercus nodules. Farmers do not bring pigs believed to be infected to the slaughterhouse for fear of confiscation. Therefore, reliable statistics on porcine cysticercosis can only be acquired at the household level. We examined the utility of the tongue test as a diagnostic tool for porcine cysticercosis. The results of the tongue test was compared with 2 serologic methods for the detection of cysticercosis, the enzyme-linked immunosorbent assay (ELISA) and the enzyme-linked immunoelectrotransfer blot assay (EITB), and with necropsy results. We examined 11 animals from an endemic area (Huancayo) and 42 animals from an area free of cysticercosis (Lima). The tongue test has a sensitivity of 70% and a specificity of 100%, the EITB a sensitivity and specificity of 100%, and the ELISA a sensitivity of 79% and a specificity of 75%. Thus, the tongue examination, being a test essentially without cost and having fair sensitivity and high specificity, can be useful in epidemiological surveys. Prevalence for porcine cysticercosis in Huancayo is 23.4% by tongue examination, 31.2% by necropsy, 37.7% by ELISA, and 51.9% by EITB.     

Comparison of enzyme-linked immunosorbent assay and indirect immunofluorescence assay in the diagnosis of human strongyloidiasis.

BACKGROUND: An enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence assay (IFA) were evaluated for serological diagnosis of human strongyloidiasis. METHODS: Serum specimens obtained from 46 individuals infected with Strongyloides stercoralis, 37 healthy persons and 381 persons with other parasitic infections were tested using an IgG-ELISA that used crude antigen of S. stercoralis filariform larvae and an IFA. Test sera were pre-incubated with antigens from Ascaris, Toxocara and hydatid protoscolices to remove non-specific antibodies. RESULTS: The sensitivity, specificity, positive predictive value and negative predictive value for ELISA were 93.5%, 96.1%, 72.9% and 99.2%, respectively, and those for IFA were 87%, 90.1%, 49.4% and 98.4%, respectively. Both assays showed false positivity in hydatidosis, ascariasis and toxocariasis; however, this was less common with ELISA. CONCLUSION: ELISA method using filariform larval antigen may be a sensitive and specific test for human strongyloidiasis, and may be preferable to IFA.     

Serodiagnosis of neurocysticercosis using synthetic 8-kD proteins: comparison of assay formats

The assay of choice for serological detection of cysticercosis in humans and pigs is the enzyme-linked immunoelectrotransfer blot (EITB), a Western blot assay that relies on the use of seven lentil-lectin-purified glycoproteins (LLGPs) derived from Taenia solium metacestodes. The EITB is has a sensitivity of 98% and a specificity of 100% in detecting cysticercosis, yet scarcity of native source material and the labor-intensive process of metacestode purification hinder its practicality. These limitations have necessitated the reproduction of the EITB antigens in synthetic forms. Four chemically synthesized LLGP antigens, TS14, TS18var1, TSRS1, and TSRS2var1, were assayed individually by enzyme-linked immunosorbent assay (ELISA) and Western blot for immunoreactivity against a large cohort of sera from clinically defined neurocysticercosis patients. The sensitivity and specificity of all four of these antigens using the ELISA format were well below the standards set by the LLGP EITB, whereas results of the Western blot format closely mirrored those of the LLGP EITB.     

Diffusion-in-gel enzyme-linked immunosorbent assay, ELISA and IFA test in the detection of malarial antibodies

Malarial antibodies in 80 patients were measured using the diffusion-in-gel enzyme linked immunosorbent assay (DIG-ELISA), enzyme-linked immunosorbent assay (ELISA) and the indirect fluorescent antibody (IFA) test. Good correlations were obtained between all three tests in terms of sensitivity and reliability. DIG-ELISA has the advantage of being a rapid diagnostic tool for the detection of malarial antibodies.     

IEST、IFAT和Dot-ELISA诊断华枝睾吸虫病的比较研究

用成虫冰冻切片抗原免疫酶染色试验(IEST)、间接免疫荧光抗体试验(IFAT)和斑点-ELISA(Dot-ELISA)对比检测了51例华枝睾吸虫病人血清,阳性率分别为92%、88%和92%.检测健康人血清50例,假阳性率分别为2%、4%和2%.用3法检测22例急性血吸虫病人血清、20例慢性血吸虫病人血清和15例肺吸虫感染者血清.IEST的交叉反应率分别为14%、5%和0;IFAT的交叉反应率分别为14%、10%和0;Dot-ELISA的交叉反应率分别为14%、5%和0.显示3种方法诊断华枝睾吸虫病均有较高的敏感性和特异性.而IEST和Dot-ELISA更适于现场应用.     

An Evaluation Study of Enzyme-Linked Immunosorbent Assay (ELISA) Using Recombinant Protein Pap31 for Detection of Antibody against Bartonella bacilliformis Infection among the Peruvian Population

Reliable laboratory testing is of great importance to detect Bartonella bacilliformis infection. We evaluated the sensitivity and specificity of the enzyme-linked immunosorbent assay (ELISA) using recombinant protein Pap31 (rPap31) for the detection of antibodies against B. bacilliformis as compared with immunofluorescent assay (IFA). Of the 302 sera collected between 1997 and 2000 among an at-risk Peruvian population, 103 and 34 samples tested positive for IFA-immunoglobulin G (IgG) and IFA-IgM, respectively. By using Youden''s index, the cutoff values of ELISA-IgG at 0.915 gave a sensitivity of 84.5% and specificity of 94%. The cutoff values of ELISA-IgM at 0.634 gave a sensitivity of 88.2% and specificity of 85.1%. Using latent class analysis, estimates of sensitivity and specificity of almost all the assays were slightly higher than those of a conventional method of calculation. The test is proved beneficial for discriminating between infected and non-infected individuals with the advantage of low-cost and high-throughput capability.     

Comparison of the enzyme-linked immunosorbent assay (ELISA) with standard tests used to detect yellow fever virus antibodies

The enzyme-linked immunosorbent assay (ELISA) was used to detect antibodies to yellow fever virus in 110 sera from patients living in an epidemic yellow fever area. The results were then compared with those obtained with the hemagglutination-inhibition (HI), complement-fixation (CF), neutralization (NT), and indirect immunofluorescence (IFA) tests. This ELISA, which used a type-specific antigen, showed the same results as the NT test and was found to be more sensitive and more specific than the HI and CF tests.