弓形虫B1基因套式PCR检测技术研究

DETECTION OF B 1 GENE OF TOXOPLASMA WITH NESTED POLYMERASE CHAIN REACTION

本文以弓形虫多拷贝的Ｂ１基因为靶基因建立了套式ＰＣＲ检测技术。弓形虫ＲＨ株、新疆、上海、昆山分离株ＤＮＡ均能出现清晰条带，而人组织及其它微生物ＤＮＡ不出现任何条带，弓形虫套式ＰＣＲ较普遍ＰＣＲ敏感１００倍。昆山分离株经套式ＰＣＲ扩增并进行ＤＮＡ测序与ＲＨ株完全一致。８０例孕妇外周血经弓形虫套式ＰＣＲ扩增４例阳性，阳性率５％。

Aim To establish a molecular diagnosis technique of toxoplasmosis Method The nested polymerase chain reaction(PCR)detected specific sequence of B 1 gene of toxoplasma Results The toxoplasma strains from Xinjiang,Shanghai,Kunshan isolated and RH ware amplified positive band by the nested PCR None of homan tissue and other organisms ware amplified The sensitivity of the nested PCR is higher 100 times than conventional PCR The direct sequencing of amplification products from Kunshan strain is quite same as RH strain The positive rate was 5% in 80 cases of pregnancy women Conclusion The nested PCR of toxoplasma is a sensitive,specific and rapid diogrosis method