中国寄生虫学与寄生虫病杂志 ›› 2000, Vol. 18 ›› Issue (5): 6-281.

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斯氏狸殖吸虫抗原分析和血清学诊断方法的研究

张锡林,段建华,王英,况明书,黄复生
  

  1. 第三军医大学基础部寄生虫学教研室,重庆 400038
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2000-10-30 发布日期:2000-10-30

ANALYSIS OF PAGUMOGONIMUS SKRJABINI ANTIGEN AND ITS APPLICATION IN SERODIAGNOSIS

ZHANG Xi lin;DUAN Jian hua;WANG Ying;KUANG Ming shu;HUANG Fu sheng   

  1. Department of Parasitology;Third Military Medical University;Chongqing 400038
  • Received:1900-01-01 Revised:1900-01-01 Online:2000-10-30 Published:2000-10-30

摘要:   [目的 ]分析斯氏狸殖吸虫囊蚴、童虫和成虫各期抗原和建立特异性抗原的斯氏狸殖吸虫病血清学诊断方法。 [方法 ]用SDS PAGE分离斯氏狸殖吸虫的各虫期抗原 ,经免疫印迹识别成虫期特异性诊断抗原。采用电洗脱技术分离 10~ 30kDa蛋白组分 ,建立纯化抗原的dot ELISA血清学诊断斯氏狸殖吸虫病。 [结果 ]斯氏狸殖吸虫病患者血清与成虫抗原的 10~ 30kDa显示较多免疫识别带 ,主带为 2 2、2 4和 2 6kDa。与血吸虫病和华支睾吸虫病患者血清的交叉反应带出现于 6 0~ 90kDa。斯氏狸殖吸虫成虫 10~ 30kDa抗原的dot ELISA与成虫粗抗原的ELISA检测 2 8例疑诊病人血清 ,两法阳性率间差异无显著性。而检测 38例感染其它吸虫和肺部疾病患者血清 ,粗抗原的ELISA交叉反应率为 13 2 % (5 /38)。 [结论 ]斯氏狸殖吸虫成虫 10~ 30kDa抗原的dot ELISA为斯氏狸殖吸虫病高度特异和敏感的血清学诊断方法。

关键词: 斯氏狸殖吸虫, 抗原, 免疫印迹, 血清学诊断

Abstract:  Objective] To analyse the soluble antigens of different developmental stages of Pagumogonimus skrjabini and deve lop a specific and sensitive serodiagnostic method for pagumogonimiasis. [Methods] The soluble antigens of P.skrjabini of various stages were separated by SDS PAGE. The specific antigen of the adult fluke was recognized immunologically by immunoblot assay. The protein bands between 10~30 kDa purified by SDS PAGE and electrophoretic elution were used in dot ELISA. [Results] Using dot ELISA, the soluble antigens of adult were recognized by sera infected with P skrjabini . More reactive bands appeared at 10~30 kDa, but major protein bands were at 22、24 and 26 kDa. However, using sera from patients infected with other trematodes including schistosome and Clonorchis , cross reaction bands appeared within 60 to 90 kDa. When compared with ELISA of crude adult antigens for detecting 28 suspected patients, there was no significant difference between the two methods. The sera of 38 patients with other diseases were also detected by the two tests. No cross reaction occurred with the purified adult antigen dot ELISA while 13 2%(5/38) of the sera cross reacted in ELISA of crude adult antigens. [Conclusion] Dot ELISA using 10~30 kDa antigen might be a specific and sensitive serodiagnostic method for diagnoing pagumogonimiasis.

Key words: Pagumogonimus skrjabini, antigen, immunoblot, serodiagnosis