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首次建立弓形虫人株(ZS_2株)基因组文库,筛选出一个弓形虫特异DNA片段的克隆。对该克隆的DNA片段(1.1kb)进行了限制性内切酶图谱分析。应用Southern印迹法及斑点印迹法检测,结果示同位素~(32)P标记的该克隆DNA片段能与弓形虫DNA、人工感染弓形虫幼猪白细胞和胸腺DNA、弓形虫感染病人DNA杂交,但不与正常人、正常幼猪外周血白细胞、正常小鼠脾脏、恶性疟原虫、肺孢子虫、pBR322的DNA杂交。斑点杂交检测低限为100个弓形虫或500pg弓形虫DNA。该探针已成功地应用于多种弓形虫感染病例的检测,为弓形虫病提供了一个特异、灵敏的DNA诊断方法。
The Toxoplasma gondii strain (ZS 2 strain) genomic library was first constructed and a clone of Toxoplasma gondii-specific DNA fragment was screened out. The cloned DNA fragment (1.1 kb) was subjected to restriction endonuclease mapping. The results of Southern blotting and dot blot showed that the ~ (32) P-labeled DNA fragment could be hybridized with Toxoplasma gondii DNA, Toxoplasma gondii leucocyte and thymus DNA, DNA from Toxoplasma gondii infection but not Normal human, normal porcine peripheral blood leukocytes, normal mouse spleen, Plasmodium falciparum, Pneumocystis, pBR322 DNA hybridization. Dot blot hybridization detection limit of 100 Toxoplasma gondii or 500pg DNA. The probe has been successfully used in a variety of cases of Toxoplasma gondii detection, for toxoplasmosis provides a specific and sensitive method of DNA diagnosis.