中国寄生虫学与寄生虫病杂志 ›› 1990, Vol. 8 ›› Issue (3): 165-169.

• • 上一篇    下一篇

弓形虫基因组文库的建立、特异克隆的筛选和弓形虫病的DNA诊断

夏爱娣 ,顾允中 ,徐世杰 ,陈诗书 ,杨惠珍,徐克继,钱宗立   

  1. 上海第二医科大学生物化学教研室; 上海第二医科大学分子生物学实验室; 上海第二医科大学寄生虫学教研室; 上海第二医科大学寄生虫学教研室; 上海第二医科大学寄生虫学教研室
  • 出版日期:1990-08-31 发布日期:2017-01-09
  • 基金资助:
    国家自然科学基金 上海市高等教育局提供资助

CONSTRUCTION OF A GENOMIC DNA LIBRARY OF TOXOPLASMA GONDII (ZS2 STRAIN), SCREENING OF SPECIFIC CLONE AND DNA DIAGNOSIS OF TOXOPLASMOSIS

  • Online:1990-08-31 Published:2017-01-09

摘要: 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。

关键词: 弓形虫, 基因组文库, 克隆, 弓形虫病

Abstract: We have constracted a genomic DNA library of Toxoplasma gondii (ZS2 strain) and screened out a specific DNA sequence for T. gondii. The restriction map of the cloned DNA fragment (1.1kb) was analysed. The Southern and dot-blot analyses showed that the 32P-labeled cloned DNA fragment hybridized to the parasite DNA, DNAs from peripheral white blood cells and thymus of baby pigs artificially infected with T. gondii and DNAs of T. gondii- positive anencephalus and hydrocephalus, but did not hybridize to DNAs from controls, i. e., normal human and baby pig peripheral white blood cells, spleen of normal mouse, Plasmodium falciparum, Pneumocystis carinii and pBR322. As few as 100 T. gondii parasites or 500pg purified DNA from T. gondii can be detected by dot blot hybridization. This established DNA probe method was specific and sensitive and has been successfully used in detecting various cases infected with T. gondii.