中国寄生虫学与寄生虫病杂志 ›› 1998, Vol. 16 ›› Issue (1): 3-15.

• 论著 • 上一篇    下一篇

PCR-ELISA检测疟原虫DNA的研究

张龙兴; 汤林华; 冯晓平; 王聚君;   

  1. 中国预防医学科学院寄生虫病研究所;
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:1998-02-28 发布日期:1998-02-28

STUDY ON DETECTION OF MALARIA PARASITE DNA BY PCR ELISA *

Zhang Longxing **; Tang Linhua; Feng Xiaoping; Wang Jujun

  

  1. Institute of Parasitic Diseases; Chinese Academy of Preventive Medicine; Shanghai 200025;
  • Received:1900-01-01 Revised:1900-01-01 Online:1998-02-28 Published:1998-02-28

摘要: 目的:介绍一种诊断疟疾的新方法PCR-ELISA。方法:根据业已报道的疟原虫SSUr-RNA基因保守序列,设计并合成一对通用于恶性疟原虫和间日疟原虫的引物,其中一引物的5’端加以生物素标记。经PCR扩增后,携带有生物素的扩增产物与先期包被于ELISA板上的亲和素结合,再经与恶性疟原虫、间日疟原虫特异、荧光素标记的寡核苷酸探针分别杂交,底物显色等步骤,使PCR产物得以半定量地检出。结果:对于恶性疟原虫和间日疟原虫,本法检出最低原虫密度阈值分别为4和10个原虫/μl血(取血20μl),本法检测两种疟原虫未发现交叉反应。结论:本试验具有较高的敏感度和特异性,可望用于疟疾流行病学调查

关键词: PCR-ELISA, 疟原虫, DNA检测

Abstract: AIM: To present a new malaria diagnostic method based on detection of malaria parasite DNA by PCR ELISA. METHODS: According to the conserved sequence of Plasmodium SSUrRNA genes reported, a pair of primers in which one primer was biotinylated and another was unbiotinylated, suitable for DNA amplification of both falciparum and vivax malaria parasites were designed and synthesized. After denaturation and washing, the incoporated biotinylated product with avidin coated on plates previously was hyb...

Key words: PCR ELISA, malaria parasite, DNA detection