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斑点免疫金过滤法(DIGFA)用于使用纯化的31-kDa抗原快速检测抗大鼠肺线虫广州管圆线虫(线虫纲:后圆线虫科)的特异性抗体。

Dot immunogold filtration assay (DIGFA) for the rapid detection of specific antibodies against the rat lungworm Angiostrongylus cantonensis (Nematoda: Metastrongyloidea) using purified 31-kDa antigen.

作者信息

Eamsobhana P, Gan X X, Ma A, Wang Y, Wanachiwanawin D, Yong H S

机构信息

Department of Parasitology, Faculty of Medicine Siriraj Hospital,Mahidol University,Bangkok10700,Thailand.

Institute of Parasitic Diseases, Zhejiang Academy of Medical Sciences,Hangzhou310013,PR China.

出版信息

J Helminthol. 2014 Dec;88(4):396-401. doi: 10.1017/S0022149X13000321. Epub 2013 May 28.

Abstract

A rapid dot immunogold filtration assay (DIGFA) was adopted for specific immunodiagnosis of human cerebral angiostrongyliasis, using purified 31-kDa glycoprotein specific to Angiostrongylus cantonensis as diagnostic antigen and protein A colloidal gold conjugate as antigen-antibody detector. A total of 59 serum samples were assayed - 11 samples from clinically diagnosed patients with detectable A. cantonensis-specific antibody in immunoblotting; 23 samples from patients with other related parasitic diseases, i.e. gnathostomiasis (n= 8), cysticercosis (n= 5), toxocariasis (n= 2), filariasis (n= 4), paragonimiasis (n= 2) and malaria (n= 2); and 25 samples from normal healthy subjects. The sensitivity and specificity of DIGFA to detect anti-A. cantonensis specific antibodies in serologically confirmed angiostrongyliasis cases, were both 100%. No positive DIGFA was observed in cases with other parasitic diseases, and the healthy control subjects. The 3-min DIGFA is as sensitive and specific as the 3-h immunoblot test in angiostrongyliasis confirmed cases that revealed a 31-kDa reactive band. The gold-based DIGFA is more rapid and easier to perform than the traditional enzyme-linked immunosorbent assay (ELISA). The test utilizing purified A. cantonensis antigen is reliable and reproducible for specific immunodiagnosis of human infection with A. cantonensis - thus can be applied as an additional routine test for clinical diagnostic support. Large-scale sero-epidemiological studies in endemic communities in north-east Thailand are under way to evaluate its usefulness under field conditions.

摘要

采用快速斑点免疫金渗滤法(DIGFA)对人体广州管圆线虫病进行特异性免疫诊断,以纯化的广州管圆线虫特异性31-kDa糖蛋白作为诊断抗原,蛋白A胶体金复合物作为抗原抗体检测剂。共检测了59份血清样本——11份来自临床诊断为广州管圆线虫病且免疫印迹法可检测到广州管圆线虫特异性抗体的患者;23份来自患有其他相关寄生虫病的患者,即颚口线虫病(n = 8)、囊尾蚴病(n = 5)、弓蛔虫病(n = 2)、丝虫病(n = 4)、肺吸虫病(n = 2)和疟疾(n = 2);以及25份来自正常健康受试者的样本。DIGFA检测血清学确诊的广州管圆线虫病病例中抗广州管圆线虫特异性抗体的敏感性和特异性均为100%。在患有其他寄生虫病的病例和健康对照受试者中未观察到DIGFA阳性。在确诊为广州管圆线虫病且显示出31-kDa反应带的病例中,3分钟的DIGFA与3小时的免疫印迹试验一样敏感和特异。基于金的DIGFA比传统的酶联免疫吸附测定(ELISA)更快且更易于操作。利用纯化的广州管圆线虫抗原进行的检测对于人体感染广州管圆线虫的特异性免疫诊断是可靠且可重复的——因此可作为临床诊断支持的附加常规检测应用。泰国东北部流行社区正在进行大规模血清流行病学研究,以评估其在现场条件下的实用性。

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