随机引物聚合酶链反应技术鉴别弓形虫株的研究

STUDIES ON THE IDENTIFICATION OF STRAINS OF TOXOPLASMA GONDII BY RANDOMLY PRIMED POLYMERASE CHAIN REACTION

本文运用随机引物聚合酶链反应技术鉴别弓形虫株，获得较满意结果。实验根据Ｍ１３噬菌体已知部分核酸序列，自行设计并合成了３条引物，在一定条件下扩增ＲＨ，ＺＳ１、ＺＳ２、ＳＨ－１等４株弓形虫基因组ＤＮＡ，扩增产物经２％琼脂糖凝胶电泳分带。扩增带型显示４株受检的弓形虫株具有株间差异。

Randomly primed polymerase chain reaction (RP-PCR) was applied to identify four strains of Toxoplasma gondii different in virulence in this article. No prior sequence of Toxoplasma gondii is required.Three random primers (RP) were designed according to the sequence of M(13) phage plaques. One of them was a consensus (universal) primer of M(13) named RP1(5'GACGGGCCAGT3'). Tel others were RP2(5'TATGACCATG3') and RP3(5'GCAACGCAAT3') respectively. The results demonstrated that different primers used to amplify the genomic DNA of four strains of Toxoplasma gondii had showed different banding patterns, the polymorphisms in genomic ″fingerprints″. By comparing the polymorphisms, four strains of Toxoplasma gondii can be distinguished.