并殖吸虫病感染血清识别的斯氏狸殖吸虫成虫抗原及其特性分析

目的 通过并殖吸虫病患者血清识别斯氏狸殖吸虫成虫抗原,研究特异性抗原的生物学特性,为特异性诊断抗原和疫苗的开发研究提供依据. 方法 成虫可溶性抗原经SDS-PAGE、2-DE、感染血清的免疫印迹识别特异性的蛋白,从识别的蛋白点中选取A、B、C 3个点做进一步分析,采用串联质谱进行蛋白肽段测序,结合免疫金标技术抗原定位,分析感染血清识别蛋白的特性. 结果 A点蛋白与6种并殖吸虫相关蛋白匹配,其中与卫氏并殖吸虫的一新型蛋白westerpain-1/10匹配分值最高,B点蛋白与5种并殖吸虫相关蛋白匹配,其中与卫氏并殖吸虫的半胱氨酸蚩白酶匹配值最高;C点蛋白无相关蛋白与之相匹配.免疫金标记技术显示感染血清识别的虫体抗原位于虫体表膜和肠腔及肠腔内容物. 结论 并殖吸虫病感染血清可识别经SDS-PAGE、2-DE分离的斯氏狸殖吸虫成虫虫体可溶性抗原.串联质谱分析以及免疫金标记检测显示其主要成分为虫体表膜与分泌排泄物.

Adult antigens of Pagumogonimus skrjabini identified by the serum with paragonimiasis and analysis on their characterizations

Objective To analyze the adult soluble antigens of Pagumogonimus skrjabini and investigate their biological characteristics. Methods The soluble antigens of adult worm of P. skrjabini were analyzed by SDS-PAGE, two-dimensional gel electrophoresis (2-DE) , Western blot and immunogold labeling technique. Results There were 3 proteins points (A,B,C) identified by tandem mass spectrometry. The A point matched 6 kinds of protein related to lung fluke, among which having the highest matching score to a new type of protein westerpain-1/10 of Paragonimus westermani. The B point matched 5 kinds of protein related to lung fluke, among which having the highest matching score to cysteine proteinase of Paragonimus westermani. The C point did not match any protein related to lung fluke. Moreover antigens of adult worm were located on the adult worm membrane and intestinal tract, indicated by immunogold labeling technique. Conclusion The main components of identified proteins analyzed by tandem mass spectrometry and immunogold labeling technique were located on membrane and in the intestinal tract of the worm.