肺吸虫和血吸虫总RNA的提取及电泳分析

Paragonimus Westermani RNA Extraction and Electrophoretic Analysis

目的为构建肺吸虫ｃＤＮＡ文库，获得目的重组抗原，用以肺吸虫病的诊断，而制备卫氏肺吸虫的总ＲＮＡ。方法采用改进的一步法（Ａ和Ｂ）提取了卫氏肺吸虫成虫的总ＲＮＡ，同时提取了日本血吸虫成虫及其虫卵、小鼠肝脏的总ＲＮＡ。结果人睾丸组织的总ＲＮＡ经琼脂糖凝胶电泳，观察到方法Ａ所提取的肺吸虫、血吸虫及其虫卵的总ＲＮＡ在凝胶上只有一条略大于１８Ｓ的条带。用方法Ｂ提取的肺吸虫以及血吸虫的总ＲＮＡ则出现２条条带，其中１条色淡，并略小于２８Ｓ，另１条色深，并略大于１８Ｓ。结论肺吸虫的Ｌ－ｒＲＮＡ中也存在切口现象。

Objective\ To construct cDNA library of Paragonimus with recombinant target antigens used to make the total RNA of Paragonimus westermani. Methods\ Total RNA was extracted from adult worms of Paragonimus westermani (P.w) and Schistosoma japonicum (S.j), eggs of S.j, mouse liver by single step isolation with acid guanidinium thiocyanate phenol chloroform mixture (method A). The method is simple, and time saving and provides RNA of high purity. Results\ Analysis on agarose gel electrophoresis indicated that RNA of mouse liver and human testis tissues revealed two major bands (28S and 18S respectively), RNA of P.w S.j eggs of S.j were slightly larger than 18S band. But two bands of RNA of P.w and S.j could be seen with another method (method B), one band being slightly smaller than 28S, another slightly larger than 18S. Conclusion\ The results confirmed that Paragonimus has cut phenomenon in its L rRNA.