中国寄生虫学与寄生虫病杂志 ›› 1999, Vol. 17 ›› Issue (3): 1-131.

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浸棒-夹心-胶体染料免疫试验检测血吸虫病患者循环抗原方法的建立及应用

谢艳辉;薛纯良;娄文娴
  

  1. 上海第二医科大学寄生虫学教研室 上海 200025
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:1999-06-30 发布日期:1999-06-30

ESTABLISHMENT AND APPLICATION OF DIPSTICK SANDWICH COLLOIDAL DYE IMMUNOASSAY FOR CIRCULATING ANTIGEN DETECTION IN SCHISTOSOMIASIS PATIENTS

XIE Yanhui;XUE Chunliang;LOU Wenxian   

  1. Department of Parasitology;Shanghai Second Medical University;Shanghai 200025
  • Received:1900-01-01 Revised:1900-01-01 Online:1999-06-30 Published:1999-06-30

摘要:   目的:建立检测感染宿主血清中循环抗原的简易免疫学试验方法。方法:从国产的工业染料中首次筛选出 1 种能用于免疫显色反应的淡紫色染料莲( H F R L),用 Ig M 类别抗日本血吸虫组合单抗 D H C 标记染料,并探索最适标记条件。建立浸棒夹心胶体染料免疫试验( D S D I A),检测宿主血清中日本血吸虫循环抗原。结果: D S D I A检测日本血吸虫 S E A 的最低检出极限为 5 ng/m l,分别检测急性血吸虫病患者 14 例及慢性血吸虫患者 113 例的血清,其阳性率分别为 100% 和 52.2% ,检测正常学生血清 113 例,特异性为 92.9% 。该法的敏感性及特异性与dot E L I S A 相似,且两法联合检测可提高检出率,而特异性不变,提示两法有一定的诊断互补叠加效应。标记染料室温下保存 1 w k 活性不变。结论: D S D I A 操作简便、试剂价廉,有广泛的应用潜力。

关键词: 胶体染料免疫试验, 日本血吸虫病, 循环抗原检测

Abstract:  AIM:To set up a simple immunoassay for the detection of circulating antigens in host serum. METHODS: A kind of colloidal pale purple dye (HFRL) used as a staining reagent for immunoassay was first selected from the dyes produced in China and an optimum condition for labelling the dye onto the combined IgM antibodies was explored. A dipstick sandwich colloidal dye immunoassay (DS DIA) for the detection of circulating antigens in host sera was established. RESULTS:The minimal concentration of SEA detected was 5 ng/ml by DS DIA. Serum circulating antigen detection in 14 cases of acute schistosomiasis japonica ,113 chronic cases and in 113 healthy controls revealed that the sensitivity was 100% in acute cases, 52.2% in chronic cases and the specificity was 92.9% for uninfected students.The sensitivity and specificity of DIA were similar to those of dot ELISA . A certain degree of diagnostic complementarity up to 76.1% in sera from these patients was seen when DS DIA and dot ELISA were used in parallel. CONCLUSION:DS DIA is a simple, economical and reliable method for detecting circulating antigens of Schistosoma japonicum ,having a wide potential value especially for field use.

Key words: Colloidal dye immunoassay (DIA), schistosomiasis japonica, circulating antigen detection