猪囊尾蚴特异循环免疫复合物致病作用的研究

本文对脑型和混合型猪囊尾蚴病患者脑脊液（ＣＳＦ）中特异循环免疫复合物（ＣＩＣ）进行了检测，其结果与患者ＣＴ表现及吡喹酮治疗第一个疗程过程中患者出现的高颅压情况存在一定联系。人工免疫复合物（ＩＣ）对ＢＡＬＢ／ｃ鼠作用后进行病理检查，发现脑实质和肾脏小血管均出现充血现象．部分小血管内皮细胞遭到破坏。因此认为特异循环免疫复合物可能是脑型、混合型猪囊尾蚴病患者在第一疗程中出现高颅压反应的主要因素之一。     

Evaluation of dot-ELISA for serological diagnosis of amebiasis

 We improved the dot enzyme-linked immunosorbent assay (dot-ELISA) reported by Itoh and Sato, and assessed the usefulness of this test for the diagnosis of amebiasis. The sensitivity of dot-ELISA was compared with that of plate ELISA, the indirect hemagglutination test (IHA), and the indirect fluorescent antibody test (IFA) for the diagnosis of amebiasis. Of 37 serum samples from patients with documented amebiasis, 36 (97.3%) were positive by dot-ELISA. There was consistency among the results of dot-ELISA, plate ELISA, and IFA, although the positive rate of IHA was lower than that of the others (78.4%; 29 of 37 cases were positive). The specificities of dot-ELISA and plate ELISA were assessed using a total of 68 sera, collected from 38 patients infected with seven different parasites other than Entamoeba histolytica, 10 patients showing diarrhea or liver abscess without parasitic infection, and 20 healthy individuals. The two assays showed no false-positive results. There were no differences in sensitivity and specificity between dot-ELISA and plate ELISA. However, the dot-ELISA technique seems to be more feasible for clinical application than plate ELISA techniques, because the assay does not require any specific equipment. Received: July 8, 2002 / Accepted: December 7, 2002 RID="*" ID="*" A summary of this paper was presented at the 74th General Meeting of the Japanese Association for Infectious Diseases (Fukuoka, April 2000). Acknowledgments The authors are indebted to Professor Tsutomu Takeuchi and Dr. Seiki Kobayashi, Department of Tropical Medicine and Parasitology, Keio University School of Medicine, for supplying E. histolytica antigen, and to Dr. Hiroshi Yamasaki, Department of Parasitology, Juntendo University School of Medicine, for supplying recombinant Toxocara canis antigen for this study.     

JPG3单克隆抗体Dot-ELISA用于血吸虫病疗效考核的探讨

目的评估JPG3单克隆抗体Dot-ELISA在血吸虫病疗效考核中的应用价值。方法以吡喹酮(50mg·kg-1,顿服)治疗血吸虫病人后,采用JPG3单抗Dot-ELISA方法,追踪检测治疗后半年、1年和2年病人血清中的循环抗原(CAg),并计算其阴转率。结果治疗后半年、1年和2年病人血清中CAg阴转率分别为70.63%、80.40%和84.34%,与抗体检测方法(IHA)的阴转率分别为15.38%、27.24%和49.23%,有非常明显的差异;比较不同感染度、不同性别和不同年龄组患者之间CAg的阴转率均没有显著性差异。结论JPG3单抗Dot-ELISA在血吸虫病疗效考核中有较好的应用价值。     

单克隆抗体Dot-ABC-ELISA检测包虫病人循环抗原

作者用活化生物素标记抗细粒棘球蚴抗原单克隆抗体（ＭｃＡｂ）１Ｄ８，以聚氯乙烯（ＰＶＣ）白色薄膜凹孔板为载体，建立了ＭｃＡｂ－Ｄｏｔ－ＡＢＣ－ＥＬＩＳＡ检测包虫病人循环抗原的方法。测定人源包虫囊液抗原的灵敏度为６ｎｇ／３μｌ样品．９２例确诊包虫病人血清循环抗原检出率为７６．１％（７０／９２），在抗体阳性者中循环抗原检出率为８１．６％（６２／７６），抗体阴性者中检出率为５０％（８／１６）。１００份正常人和１２２份羹虫病等其它寄生虫病人血清，仅１份囊虫病人血清出现交叉反应．测定循环抗原对包虫病人，尤其是血清抗体阴性者的免疫诊断有重要价值．本法简便、快速，适于推广．     

Saliva as a source of anti-Toxoplasma gondii IgG for enzyme immunoassay in human samples

Toxoplasmosis, a highly prevalent disease, is mainly diagnosed by serology. Incidence studies could be feasible in children, but ethical concerns restrict blood sampling in this group. Saliva contains small amounts of crevicular fluid IgG. Dot-ELISA and a protein A IgG capture immunoassay were standardized for anti-Toxoplasma gondii IgG in paired saliva and serum samples from 20 adult volunteers. A frequency of toxoplasmosis of 19% (95% CI 12–28) was found in 100 saliva samples from university graduates using both assays. Toxoplasmosis immunoassays using saliva IgG are a promising tool for the investigation of the epidemiology of this disease in children and other vulnerable groups.     

三种血清学方法对马来丝虫病监测的应用研究

目的：比较三种方法监测马来丝虫病的效果。方法：间接荧光抗体试验（ＩＦＡＴ）、酶联免疫吸附试验（ＥＬＩＳＡ）、斑点－酶联吸附试验（Ｄｏｔ－ＥＬＩＳＡ）。结果：检测１９９０、１９９１、１９９４年基本消灭马来丝虫病的流行区人群血清，ＩＦＡＴ阳性率分别为２４８５％、２０３２％和８３０％；ＥＬＩＳＡ阳性率分别为２１６６％、３０３２％和１５８４％；Ｄｏｔ－ＥＬＩＳＡ阳性率分别为３２５０％、１０９６％和１８８６％；总阳性率分别为４６６６％、３５８０％和３２０７％。结论：以ＩＦＡＴ法较为稳定，３种方法联合应用，可提高阳性率     

用双夹心斑点免疫金银染色法检测病人血中疟疾循环抗原

应用抗间日疟原虫和抗恶性疟原虫红内期抗原单克隆抗体建立了双夹心斑点免疫金银染色法和双夹心免疫酶斑点法并用于检测间日疟和恶性疟患者血清中循环抗原。共检测20份间日疟血清和15份恶性疟血清,双夹心斑点免疫金银染色法阳性率分别为90％和93％,可检出最低原虫密度为0.0009％;双夹心免疫酶斑点法阳性率分别为85％和87％,可检出最低原虫密度为0。0075％。与包虫病。弓形虫病和乙肝表面抗原阳性血清无交叉反应,检测60份健康献血员血清,两法分别有5％和8％假阳性。初步结果提示双夹心斑点免疫金银染色法检测疟疾循环抗原敏感度较高,若经过适当改进,有可能用于疟疾现场诊断和流行病学调查。     

上饶县1998年洪灾后的钩体病疫情调查

１９９８年入夏以来，在长江流域发生了历史罕见的全流域大洪水，作者参加了卫生部派往江西省灾后防病工作考察组。本文选取钩体病发病较多的江西省上饶县，进行了钩体病疫情的调查分析。上饶县于７月中旬遭受水灾，７月２９日报告首例钩体病，６～７天后发病达到高峰，２～３周后疫情得到控制。截止到９月２日，全县累积发病人数为４５６例，发病率６６.０９／１０万。其中男３５５例，女１０１例，男女构成比为３.５∶１。病死４例，男３例，女１例，病死率为０.８８％。采集３４份临床诊断为钩体病的病人血清，对临床诊断进行实验室复核，结果显示:显微镜凝集试验（ＭＡＴ）阳性率７６.４７％（２６／３４），酶免疫斑点试验（ｄｏｔ－ＥＬＩＳＡ）阳性率８２.３５％（２８／３４）。     

rSj26-Sj32-HRP-IgG-Dot-ELISA用于急性日本血吸虫病患者的诊断价值

目的探讨rSj 26-Sj 32-HRP-IgG-Dot-ELISA用于急性日本血吸虫病患者的诊断价值。方法利用纯化的rSj26-Sj32融合蛋白和日本血吸虫成虫抗原(SjAWA)建立HRP-IgG-Dot-ELISA检测急性日本血吸虫病患者血清,并以华支睾吸虫病、卫氏并殖吸虫病、棘球蚴病、乙型肝炎、肺结核病患者和健康人血清作对照,观察二者的检测结果。结果 rSj 26-Sj 32和SjAWA检测急性日本血吸虫病患者阳性检出率均为100%,特异性分别为95.35%和93.02%;二者均与华支睾吸虫病、卫氏并殖吸虫病和泡型棘球蚴病患者血清存在不同程度的交叉反应。结论纯化的rSj26-Sj32融合蛋白是一种较好的免疫诊断抗原。     

Rapid and simple detection of a mycobacterium circulating antigen in serum of pulmonary tuberculosis patients by using a monoclonal antibody and Fast-Dot-ELISA

OBJECTIVES: Several immunoassays have been established for detection of Mycobacterium tuberculosis (MTB) antigens in serum, sputum and cerebrospinal fluid of tuberculous patients using polyclonal or monoclonal antibodies raised against different mycobacterium antigens. Some of these assays display both high sensitivity and specificity for the detection of these antigens. However, these assays require special and highly expensive equipment and the procedures require long periods for their completion. Thus, the rationale of this study was to establish and evaluate Fast-Dot-Enzyme-Linked Immunosorbent Assay (FD-ELISA) as a fast, cheap and field applicable assay for detection of mycobacterium antigen in serum of patients with pulmonary TB. DESIGNS AND METHODS: This study included three groups: group I: 175 tuberculous patients with pulmonary TB proves with sputum Ziehl-Neelsen (ZN) for acid-fast bacilli and sputum culture (all cases were culture positive for MTB); Group II: 65 patients with diseases other than pulmonary TB as bronchial carcinoma (17 patients), bronchial asthma (29 patients) and chronic obstructive pulmonary disease (19 patients); group III: 50 healthy individuals. Groups II and III served as negative control groups. The target mycobacterium antigen was identified in both crude mycobacterium antigens extract and serum of patients with pulmonary TB, using western blotting technique and anti-TB monoclonal antibody (TB20-mAb) and then it was estimated in the serum samples of all studied groups as an index of tuberculosis, using a newly developed FD-ELISA. RESULTS: The target mycobacterium antigen was identified at 20 kDa molecular mass in crude mycobacterium antigens extract as well as in serum of patients with pulmonary TB. The developed FD-ELISA detected the mycobacterium antigen in the sera of 159 out of 175 pulmonary TB patients with a sensitivity of 90.8% and 93.0% positive predictive value (PPV). In addition, it identified 12 false weakly positive cases out of 115 samples of negative control groups (7 out of 65 non-TB patients and 5 out of 50 healthy individuals) with a specificity of 89.6% and 86.6% negative predictive value (NPV). Standardization of the FD-ELISA using a serial dilution of the purified mycobacterium antigen indicated that the assay was able to detect 1.8 microg/ml as a lowest detectable antigen concentration. CONCLUSIONS: The newly developed FD-ELISA is a simple, rapid and highly sensitive assay for detection of mycobacterium antigen in patients with pulmonary TB. Moreover, all steps were performed at room temperature and without the need to use expensive equipment, and this may enhance the application of this assay in tuberculosis screening programs. Further study is needed for confirmation of FD-ELISA reproducibility in light infected pulmonary TB patients and in a large population.