牛、羊、人体内寄生的细粒棘球蚴可溶性蛋白组分的SDS-PAGE分析及其分类意义评价

用十二烷基硫酸钠—聚丙烯酰胺凝胶电泳对比分析了牛、羊、人源细粒棘球蚴囊液(EgCF)及原头节(EgPS)的蛋白组分,发现三源EgCF蛋白组分相似,而三源EgPS有所差异。以羊源与人源的EgPS蛋白组分相似,牛源EgPS蛋白组分则比前二者少且图形特别。推断寄生于甘肃的黄牛、绵羊、人的细粒棘球蚴很可能是同一虫株,牛源EgPS的差异可能由于同一虫株寄生于非适宜中间宿主致发育不良所致。     

夹心ABC-ELISA检测包虫病人循环抗原的评价

本文评价了夹心ABC-ELISA对包虫病人体内循环抗原(HcAg)的检测方法。 用夹心法测HcAg,首先要制备一针对HcAg较强而特异的免疫抗体,其关键又在于用什么质量的抗原免疫动物。我们用半饱和硫酸铵盐析纯化羊源细粒棘球蚴囊液(EgCF),它去除了包括白蛋白在内的大部分宿主成分。用该抗原免疫家兔,产生兔抗细粒棘球蚴囊液抗体(RA-EgCFG)。以此种抗体对各种来源的     

ABC-ELISA技术临床诊断包虫病的研究

应用ABC-ELISA法和常规ELISA法对41例手术证实包虫病人血清及34例健康人血清检测结果表明:ABC-ELISA法是更为敏感和特异的包虫病诊断方法。该法在待测血清作1:3200稀释时无假阳性反应,阳性检出率为90.24％,而常规ELISA法假阳性率为26.5％,阳性率仅为63.47％,两法具有显著性差异(P<0.05)。此外,通过人源与羊源包虫液抗原检出结果的比较,讨论了ABC-ELISA法诊断包虫病选用异源抗原的意义。     

棘球蚴病与囊尾蚴病免疫诊断中交叉反应的动物实验观察

在包虫病和囊虫病免疫诊断中普遍存在交叉反应现象。由于病人来源差异,对较全面了解其机理带来了一定限制。为此我们分别用绵羊细粒棘球蚴囊液(EgCF)、原头节(EgPS)和猪囊尾蚴囊液(CcCF)、头节及囊壁(ccSCW)免疫小白鼠,一个月后用 EgCF 和 CcCF。致敏的血球测其抗体水平及交叉反应情况,现将结果报告如下。     

包虫和猪囊虫抗原免疫化学性质及酶联免疫印斑技术临床应用研究

用 SDS 聚丙烯酰胺凝胶电泳和酶联免疫印斑技术分别对人源细粒棘球蚴囊液、棘球蚴砂、猪囊尾蚴囊液抗原组分进行分析研究,并对两种病血清学交叉反应进行了试验。结果显示包虫囊液抗原有29条蛋白区带,有两条糖蛋白带。猪囊虫囊液抗原显示36条蛋白带谱,两条糖蛋白带。棘球蚴砂显示23条蛋白带谱。包虫病人血清与印有包虫囊液抗原的硝酸纤维膜反应显示23条反应带,其中特异带为6条。包虫病人、猪囊虫病人血清与此膜反应显示4条共同反应带。包虫病人血清与棘球蚴砂抗原膜反应显示4条主要特异反应带,包虫病人血清、猪囊虫病人血清与此膜反应显示1条共同反应带。猪囊虫病人血清与猪囊虫囊液抗原膜反应显示22条反应带,其中有5条特异反应带;包虫病人和囊虫病人血清与此膜反应显示3条共同反应带。     

不同血清学方法对调查现场包虫病人血清的检测及评价

本文应用青海囊型包虫抗原 Dot- EL ISA、IHA和新疆囊型包虫抗原 EL ISA、泡型包虫 EM18抗原 EL IB对流行病学调查现场的 2 0 6例包虫病人血清进行了检测。结果表明青海囊型包虫抗原 Dot- EL ISA和 IHA对囊型包虫病人血清的阳性率分别是 90 .37%和 91.98% ;新疆囊型包虫抗原 EL ISA对囊型包虫病人血清的阳性率为 75 .94% ;三种方法对钙化灶型囊型包虫病人血清的阳性率分别是 77.2 7%、81.82 %和 6 5 .91% ,显著低于其它囊型包虫病人血清的阳性率 ;阴性者主要是单纯性肺脏和肝脏囊型包虫病人血清。3种方法 B超、X线诊断的泡型包虫病人的血清阳性率均为 10 0 .0 %。泡型包虫抗原 EM18-EL IB检测结果表明 ,B超 ,X线诊断的泡型包虫病人血清阳性率为 73.6 8% ;囊型包虫病人血清阳性率为 5 .88% ,其中钙化灶型病人血清阳性率为 15 .91% ;阳性血清数与阴性血清数比例约为 1:7(2 5 :181)。对不同方法在囊型和泡型包虫病诊断与鉴别诊断中的价值及意义进行了讨论     

包虫与囊虫囊液中共同抗原比较

许多蠕虫存在有种属间的共同抗原成分、致使引起免疫诊断的交叉反应,包虫囊液(EgCF)与猪囊尾蚴囊液(CcCF)即含有大量的属间共同抗原。作者以间接法 ELISA 采用 EgCF     

盐析纯化包虫囊液抗原及其应用价值的评价

本文评价了半饱和硫酸铵盐析纯化包虫囊液抗原方法及其在诊断包虫病中的应用价值。虽盐析方法简单,但其提纯的包虫囊液经免疫印迹证实,去除了不少非包虫抗原性蛋白组分(由粗囊液20条带减为14条带)及保留了较多的抗原信息(剩余蛋白带大多与包虫病人血清呈阳性反应)。用两剂量抗原诊断(ELISA)40例包虫病人,盐析纯囊液比粗囊液无论是阳性符合率(1μg/ml时:85％对70％;0.3μg/ml时:75％对55％)或免疫比值(1μg/ml时:12.33±8.58对5.01±3.59;0.3μg/ml时:9.34±8.15对4.48±5.79)都有所提高。盐析纯囊液用于IHA中,也使52例正常人与46例包虫病人的凝集滴度拉开距离,阳性、阴性模式界线分明。     

包虫病诊断试验IEP和DD_5的染色前后观察

我们用琼脂凝胶免疫电泳和双向扩散试验技术对包虫病患者术前血清进行诊断分析,观察发现考马氏蓝染色前后的阳性率有很大差别,现报告如下。 包虫病人血清63例,55例系手术证实的术前血,8例为临床诊断病人(ID、IHA、ELISA均阳性)。正常人血清共20份,均为健康者血清。用羊肝包虫囊液冻干粉末做为抗原,IEP所用抗原浓度为200mg/ml,DD_5为50mg/ml。IEP和DD_5试验均按常规方法进行。     

细粒棘球蚴重组抗原B 8-kDa亚单位1对囊型包虫病的血清学诊断价值

目的通过基因工程技术获得细粒棘球蚴抗原B8-kDa亚单位1重组蛋白(rEgAgB8/1),探讨其对囊型包虫病(CE)的血清学诊断价值。方法将构建的rEgAgB8/1原核表达质粒(pET32b-rEgAgB8/1)转化至E.coli BL-21(DE3)中,用IPTG诱导表达,经亲和层析纯化获得高纯度rEgAgB8/1,以rEgAgB8/1为抗原,应用ELISA和Immuno blotting方法对31例手术确诊的囊型包虫病病人血清进行了回顾性检测与分析。结果ELISA和Immuno blotting方法检测CE病人血清阳性率均为90.3%(28/31),3例血清学检测阴性的CE病人均为初次诊断为CE及单纯性肝脏单发感染的病人;血清抗体水平随着病人棘球蚴囊数目增加而有所增加,棘球蚴囊的数目与血清抗体水平的比较用单因素方差分析有显著性差异(F=5.06,P=0.0142),1个囊与2个囊/3个囊组间血清抗体水平有显著差异,2个囊与3个囊组间差异无统计学意义。结论rEgAgB8/1重组蛋白抗原对囊型包虫病有较高的血清学诊断价值,多囊型包虫病人血清抗体水平高于单囊型包虫病人。     

应用生物素-亲和素系统提高包虫病免疫诊断的敏感性

本文对33例包虫病人的血清用ABC-ELISA法和常规ELISA法进行了检测。结果证明,ABC-ELISA法的敏感性相当于常规ELISA法的2.7倍,并有2例ABC-ELISA法为阳性,而常规ELISA法为阴性。这说明ABC-ELISA法敏感性高,并可减少常规ELISA法的假阴性率。在比较肝和肺包虫病人抗体滴度的分析中,肺包虫病人血清抗体滴度比肝包虫病人的高。     

Immunoelectrophoresis tests showing Echinococcus granulosus arc 5 in human cases of Echinococcus vogeli and cysticercosis-multiple myeloma

Human sera from one case of polycystic hydatidosis due to Echnincoccus vogeli and from a case of cysticercosis and multiple myeloma were positive to the immunoelectrophoresis (IEP) test for hydatidosis based on the E. granulosus arc 5 positivity criterion. Arc 5 can therefore be elicited in IEP tests of human sera not only by E. granulosus and E. multilocularis but also by E. vogeli antigens. Whether the cross reaction observed in the second serum was due to the multiple myeloma or to the cysticerci remains to be determined. Although arc 5 antigens are known to be present in Taenia hydatigena cyst fluid this is the first report of an arc 5 due to antigens other than Echinococcus in IEP of human sera.     

亲和素—生物素化辣根过氧化物酶复合物法诊断...

The Avidin-Biotin-Peroxidase Complex enzyme-linked immunosorbent assay (ABC-ELISA) with antigen from gerbils infected with alveolar hydatid (GAH) was performed on the sera of 54 patients with multilocular echinococcosis, 107 patients with other diseases and 102 healthy adults. All the sera were diluted 1:200 and an S/N value greater than 2.2 was taken as positive. Both the sensitivity and specificity were 98.1%. The S/N value (mean +/- SD) of sera of the patients with multilocular echinococcosis, other parasitic diseases, tuberculosis, other diseases and healthy adults were: 5.9 +/- 3.0, 1.1 +/- 0.4, 1.4 +/- 0.8, 1.1 +/- 0.5 and 1.0 +/- 0.4. The geometric mean titre (GMT) of antibody measured by ABC-ELISA was 13.7 times that of ELISA. The quantity of serum used in ELISA was 8 times that of ABC-ELISA. ABC-ELISA might be particularly helpful to identify cases with low immune responsiveness. In the detection of alveolar hydatidosis and cystic disease with GAH antigens, the positivity was 98.1 and 67.1% with 87 and 25.6% having S/N greater than 5, respectively. It is suggested that in the diagnosis of hydatid diseases by ABC-ELISA, homologous antigen should be used. The temperature and the incubation time needed for the present assay are practical, for the total time requires only 55 minutes.     

Immunological study of hydatidosis. I. Evaluation of immunoelectrodiffusion tests and enzyme-linked immunoelectrodiffusion assay (ELIEDA) in human hydatidosis.

The comparative sensitivity, specificity and rapidity of immunoelectrodiffusion (IED) on cellulose acetate membranes and of immunoelectrophoresis (IEP) on agarose gel, were evaluated in the diagnosis of hydatidosis. Pooled crude fluid from several cattle hydatis cysts was used as antigen in tests on 1,750 non-hydatis and 400 hydatis sera obtained from patients with hepatic, pulmonary, splenic, peritoneal and cerebral hydatidosis before and after surgery. Coupled to a specific human reference serum for arc 5, IED shows a specificity comparable to that of IEP but its sensitivity is slightly higher and the amount of antigen needed is very small. The appearance of a typical "gloved finger" pattern in sera from patients with ruptured cysts emphasizes the interest of quick results (3 hours) obtained by this method. In order to increase the sensitivity of IED and to define the class of immunoglobulins involved in the antigen-antibody reaction, we have coupled this method to an enzymatic technique. The immune complexes precipitated by IED were treated with peroxidase-labelled antibodies specific to each class of human immunoglobulins. The specificity of this enzyme-linked immunoelectrodiffusion assay (ELIEDA) permits one to follow the immunologic evolution of hydatidosis and to identify IgM in ruptured hepatic cysts and IgA in pulmonary cysts.     

Application of biotin-avidin system, determination of circulating immune complexes, and evaluation of antibody response in different hydatidosis patients

The avidin-biotin-peroxidase complex enzyme-linked immunosorbent assay (ABC-ELISA) and standard ELISA were used for the detection of Echinococcus granulosus antibody in sera of 101 patients operated on for hydatid disease, 40 patients with miscellaneous nonhydatid diseases, and 61 normal subjects. Sensitivity and specificity of the two procedures were comparable and the geometric mean antibody titer detected with ABC-ELISA was higher than with standard ELISA. The ABC-ELISA is a sensitive, specific, simple, and convenient method for diagnosing hydatidosis.     

用ABC—ELISA比较同种和异种抗原在诊断包虫病上的价值

用ABC-ELISA法比较研究了HHF和GAH抗原对82例手术证实包虫病患者,107例非包虫病患者和86例健康人的诊断价值。在以S/N≥2.0为阳性标准时,阳性率前者为90.24％,后者为67.07％,假阳性率各为4.08％和3.57％。抗体滴度在1:3200～6400范围者,前者占73.8％,非常显著高于后者的45.12％。说明在诊断包虫病时宜用同种抗原。本试验采用的孵育温度和时间,使孵育总时间由原来的2～4小时减少到55分钟,提高工效1～4倍,有实用价值。     

An enzyme-linked immunosorbent assay using an arc 5 antigen for the diagnosis of cystic hydatid disease

We report on an easily standardized enzyme-linked immunosorbent assay (ELISA) for the diagnosis of cystic hydatid disease. The antigen used is commercially available, and purified to elicit an arc 5 precipitation line by immunoelectrophoresis (IEP) with sera of patients harbouring hydatid cysts. The IgG-ELISA was highly sensitive and specific, and of diagnostic value compared to total Ig-ELISA, IgM-ELISA or IgA-ELISA. When compared to the indirect hemagglutination test and to counterimmunoelectrophoresis using an arc 5 antigen, minimal cross-reactions were observed in sera of patients with a presumptive diagnosis of hydatidosis but none were observed in those harbouring intestinal helminths, schistosomes or filarial parasites. The assay is of low cost, simple to perform, and highly reproducible. The IgG-ELISA provides an unequivocal laboratory diagnosis of hydatidosis. It is eminently suited for accurate and early identification of hydatid disease patients, for instance in mass surveys, who may benefit immediately from currently available anthelmintics, thereby obviating the need for surgery later.     

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.