亲和素-生物素化辣根过氧化物酶复合物法诊断泡球蚴病的研究

用沙鼠泡球蚴抗原对泡球蚴、非泡球蚴病患者和健康者进行亲和素-生物素化辣根过氧化物酶复合物的酶联免疫吸附试验(ABC-ELISA)。以样品孔血清(S)与参考阴性血清(N)的比值S/N≥2.2为阳性标准时,该法的敏感性和特异性均为98.1%。ABC-ELISA抗体几何平均滴度为ELISA的13.7倍。泡球蚴和棘球蚴病患者对该抗原反应阳性率各为98.1和67.1％,有显著性差异(P<0.01)。本试验的总孵育时间由2～4h减为55min。因此,ABC-ELISA更宜于检测微量抗体和低抗体水平患者。     

用免疫印迹法评价多房棘球蚴18kDa与16kDa抗原诊断…

目的：探讨多房棘球绦虫原头节１８ｋＤａ、１６ｋＤａ抗原（Ｅ．ｍ１８ｋＤａ和Ｅ．同６ｋＤａ）诊断泡球蚴病的应用价值。方法：应用免疫印迹法检测２４例泡球蚴病５５例囊型包虫病、３３例囊虫病和３０分健康人血清对Ｅ．ｍ．１８ｋＤａ和Ｅ．ｍ．１６ｋＤａ的抗体反应，同时用Ｅ．ｍ２－ＥＬＩＳＡ试剂盒对比检测上述血清对Ｅ．ｍ．２的抗体反应。结果：在免疫印迹试验中，Ｅ．ｍ．１８ｋＤａ和Ｅ．ｍ．１６ｋＤａ均能被所有２４     

人、沙鼠、小鼠泡球蚴抗原诊断泡球蚴病的比较研究

用同一组47例泡球蚴患者和115例对照血清比较研究了人、沙鼠和小鼠泡球蚴抗原的诊断价值。人、沙鼠和小鼠泡球蚴抗原的阳性率各为97.8％、100％和97.8％,三者无显著差异(P>0.05)。三者的假阳性率也无显著差异(P>0.05)。用沙鼠泡球蚴抗原检测时,95.74％病例的消光值在0.70以上,非常显著地高于人(48.94％)和小鼠(57.45％)抗原,P均<0.01。在1:1600以上抗体滴度组中,沙鼠泡球蚴抗原检出74.47％病例,非常显著地高于人(6.38％)和小鼠(29.79％)泡球蚴抗原(P均<0.01)。以滴度几何均数计,沙鼠泡球蚴抗原检测最高为1:1209,是小鼠泡球蚴检测的2.89倍和人泡球蚴抗原检测的3.32倍。说明在诊断人体泡球蚴病时宜用沙鼠泡球蚴抗原。以SDS-PAGE分离的人、沙鼠和小鼠泡球蚴抗原的蛋白质带数各为23、36和15。讨论了蛋白质带数与泡球蚴病诊断的关系。     

泡球蚴病和棘球蚴病患者血清IgG、IgA和IgM水平的比较

比较测定了91例泡球蚴病患者和83例棘球蚴病患者血清中IgG、IgA和IgM水平。泡球蚴病患者IgG、IgA和IgM水平及棘球蚴病患者IgG、IgM水平均非常显著地高于健康人。泡球蚴病患者IgG和IgM水平的几何均数各为棘球蚴病患者的1.67和1.25倍。IgG浓度在200IU/ml以上者,泡球蚴病患者非常显著地高于棘球蚴病患者,而IgM浓度在100IU/ml以上者,前者也显著地高于后者。用PPA-ELISA和ABC-ELISA检测的抗体效价中,只有IgG浓度和ELISA抗体效价间有显著相关(0.05>P>0.02)。     

抗辣根过氧化物酶和猪囊尾蚴的双特异单克隆抗体的研制

抗辣根过氧化物酶和猪囊尾蚴的双特异单克隆抗体的研制李达，沈喜，任永卫双特异单克隆抗体（Ｂｉｓｐｅｃｉｆｉｃｍｏｎｏｃｌｏｎａｌａｎｔｉｂｏｄｉｅｓ）是采用杂交瘤技术、化学偶联或基因工程方法制备的能够识别两种抗原的抗体或抗体复合物。近年来双特异单克隆抗...     

Evaluation of a coproantigen enzyme-linked immunosorbent assay for the diagnosis of canine echinococcosis in Iran

Summary Echinococcosis is one of the most important zoonosis in Iran. Due to this fact that providing a reliable diagnostic method for detection of this infection in definitive host is a critical prerequirement for the establishment of appropriate control programs in our country, one hundred and sixteen carnivores including 80 dogs, 27 jackals, 8 foxes and one wolf were collected from rural areas of Hamadan, Azarbaijan and Tehran provinces and examined for Echinococcus granulosus infection. Canine echinococcosis was diagnosed upon direct microscopic examination of intestinal contents and mucosal scraping for adult tapeworms, and a coproantigen detection enzyme linked immunosorbent assay (CA-ELISA) for Echinococcus granulosus. The overall prevalence of canine echinococcosis using the ELISA test was 43.1 % (50/116). The relative frequency of canine echinococcosis was 37 % (43/116) by microscopic examination. The sensitivity and specificity of the CA-ELISA test as referenced by necropsy findings was 72.1 % and 74 % respectively. We found this assay to be a very suitable and advantageous method for the surveillance of canine population especially in regions with endemic echinococcosis.     

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.     

An enzyme-linked immunosorbent assay for detection of IgG1 antibodies specific to human cystic echinococcosis in Egypt

Human cystic hydatidosis (cystic echinococcosis) is a chronic zoonotic disease that results from infection with the dog tapeworm Echinococcus granulosus. In Egypt, cystic echinococcosis (CE) is recognized in slaughtered livestock by veterinarians, however, there is little information about human CE infection rates. We describe an immunological assay useful for the diagnosis of human cystic hydatidosis. Sera were collected from surgically confirmed hydatid cases (34), nonendemic subjects free from parasitic infection (20) and from subjects (109) infected with other helminths (Hymenolepis nana, Schistosoma mansoni, Fasciola hepatica and Ancylostoma duodenale). Hydatid cyst fluid (HCF) of camel origin was used as antigen in an ELISA format to measure total E. granulosus specific IgG antibodies and IgG subclasses. Sensitivity measurements of total IgG, and IgG1-4 were 100, 100, 79.4, 61.8 and 55.9%, respectively, whereas respective specificity reached 65.1, 97.7, 98.4, 96.1 and 83. 7%. The diagnostic value of measuring IgG1 (97.7%), as assessed by a rating index (J) for combined sensitivity and specificity, was superior to total IgG (65.1%) and IgG2-4 (77.8, 57.9 and 39.6%, respectively). These findings set the stage for field evaluation of the IgG1 assay in areas endemic with human cystic hydatidosis.     

Determination of thrombin-hirudin complex in plasma with an enzyme-linked immunosorbent assay.

An enzyme-linked immunosorbent assay (ELISA) was developed for the determination of the thrombin-hirudin complex (TH) in plasma. The test is based on the sandwich principle and uses appropriate antibodies which selectively bind the corresponding moieties of the complex. The assay was calibrated by adding performed TH to normal human citrated plasma. The detection limit of the assay was 1.2 ng/ml. Mean coefficients of variation of 6.0% (intra-assay) and 6.4% (inter-assay) were found. The presence of TH was demonstrated in normal human plasma that was spiked with hirudin and subsequently activated in vitro by calcium-thromboplastin to generate thrombin. This complex was also found in plasma samples from pigs which had been treated with hirudin during experimentally induced septicaemia.     

Utilization of a peroxidase antiperoxidase complex in an enzyme-linked immunosorbent assay of elastin-derived peptides in human plasma

Chronic obstructive pulmonary disease (COPD), a major cause of morbidity and death in the smoking population, develops insidiously over many years, and significant impairment of lung function usually occurs before the disease is diagnosed. Because lung elastin degradation appears to be a prerequisite for the development of the disease, immunologic detection of elastin-derived peptides in the blood might be an effective approach to the early detection and monitoring of the disease. We here report an improved enzyme-linked immunosorbent assay for elastin peptides using a peroxidase-antiperoxidase complex as the reporter group. The assay is sensitive to 2 ng/ml elastin peptides. We show that for optimal, reproducible results the assay should be carried out at 16 degrees C rather than at room temperature and that determinations should be made on plasma containing protease inhibitors rather than on serum. The levels of elastin-derived peptides appeared to remain relatively constant when multiple samples were taken during a 5- to 10-wk period from individual subjects. In addition, patients with COPD had elevated elastin peptide levels (127 +/- 47 ng/ml) compared with levels in normal nonsmokers (58 +/- 17 ng/ml), whereas normal smokers had values intermediate between the 2 groups (mean peptide levels of 76 +/- 42 ng/ml). A small group of normal smokers (20%) had elevated elastin peptide levels similar to those in the emphysema group and may represent that group of smokers who are at risk of developing obstructive lung disease.     

结核γ-干扰素酶联免疫斑点检测在结核性心包积液诊断中的应用

目的 探讨结核菌特异性γ-干扰素（interferon-gamma,IFN-γ）酶联免疫斑点（enzyme-linked immunosorbent spot,ELISPOT）检测对结核性心包积液的诊断价值.方法 采用结核菌特异性IFN-γ ELISPOT技术同时检测20例结核性心包积液患者（TP组）和14例非结核性心包积液患者（Non-TP组）外周血单核细胞（peripheral blood mononuclear cells,PBMC）及心包积液单核细胞（pleural effusion mononuclear cells,PEMC）中结核菌特异性IFN-γ水平.结果 TP组PBMC和PEMC结核菌特异性IFN-γ水平均显著高于Non-TP组,差异有统计学意义（P＜0.01,P＜0.01）.TP组心包积液中结核菌特异性IFN-y水平显著高于外周血IFN-γ水平,差异有统计学意义（P＜0.01,P＜0.01）.PBMC ELISPOT检测结核性心包积液的敏感性和特异性分别为80.0％和85.7％;而PEMC ELISPOT敏感性和特异性为90.0％和85.7％.结论 结核菌特异性IFN-γELISPOT技术对结核性心包积液诊断和鉴别诊断具有很好的辅助价值.     

Serologic diagnosis of bone and joint tuberculosis by an enzyme-linked immunosorbent assay

Sera from patients receiving treatment for active bone and joint tuberculosis and sera from patients with inactive bone and joint tuberculosis were examined by an enzyme-linked immunosorbent assay for antibody to antigen 6, a homogeneous protein prepared from Mycobacterium tuberculosis strain H37Ra by immunosorbent affinity chromatography. Sera from 21 control subjects had a geometric mean titer of 1:6 with no difference between tuberculin purified protein derivative-positive and -negative patients. Sera from 20 patients with inactive disease had a geometric mean titer of 1:19. Fifteen patients receiving treatment for M. tuberculosis infection had a geometric mean titer of 1:179, which is significantly different from the geometric mean titers of both of the patients with inactive tuberculosis (P less than 0.001) and the control subjects (P less than 0.001). At a cut-off titer of 1:32, the sensitivity of the assay is 94% and the specificity for the control subjects and patients with inactive disease was 100%.     

Immunodiagnosis of human fascioliasis by enzyme-linked immunosorbent assay using excretory-secretory products

In sera from patients with fascioliasis the enzyme-linked immunosorbent assay (ELISA) was used to detect antibody using excretory-secretory products (ES) from Fasciola hepatica adult worms. The specificity of ES-ELISA (with OD values greater than 0.38) allowed the differentiation among fascioliasis, schistosomiasis, clonorchiasis, and other human parasite infections.     

Microtiter enzyme-linked immunosorbent assay for diphtheria antitoxin in human sera--application of parallel line assay method

The antibodies against Diphtheria toxin in human sera were measured by the ELISA method and the antitoxin titer was calculated using the parallel line assay method which is a quantitative bioassay method. The reproducibility of this method and the comparison of the titer between this method and cell culture method were proven. The dose response curve of standard antiserum and test serum calculated by the parallel line assay method showed linearity, and the regression line of the test serum was parallel to the standard serum. The coefficients of variance (CV) of the antitoxin titers obtained from triplicate measurements of 3 serum samples ranged from 9.1 to 36.0%. The serum at low titer gave a higher CV level. A good correlation was observed between the Diphtheria antitoxin titer by the neutralizing test in cultured cells and that measured by ELISA by the parallel line assay method. The coefficient of regression was 0.996 and the coefficient of the correlation was 0.899.