中国寄生虫学与寄生虫病杂志 ›› 1989, Vol. 7 ›› Issue (3): 174-176.

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恶性疟原虫DNA探针检测血内恶性疟原虫的初步研究

杨健良,杨仲炎,罗国湘,王祥荣,李菊林,杨存性   

  1. 江苏省寄生虫病防治研究所
  • 收稿日期:2017-01-09 修回日期:2017-01-09 出版日期:1989-08-31 发布日期:2017-01-09

A PRELIMINARY STUDY ON DETECTING PLASMODIUM FALCIPARUM IN BLOOD BY PLASMODIUM FALCIPARUM DNA PROBE

  • Received:2017-01-09 Revised:2017-01-09 Online:1989-08-31 Published:2017-01-09

摘要: 从培养恶性疟原虫(Fcc-1)中分离纯化基因组DNA,用~(32)P标记,作为探针,按DNA斑点杂交法,检测血样。结果表明:该探针可检出9个恶性疟原虫感染红细胞/10_6红细胞;在现场应用时,与13例恶性疟阳性标本镜检符合率为92%:与1例间日疟原虫病例和正常人血、白细胞之间,未发现非特异性杂交。

关键词: 恶性疟原虫, DNA探针检测, 基因组DNA, 防治研究, 寄生虫病, 江苏省, 放射性强度, 疟原虫感染红细胞, 杂交反应, DNA斑点杂交

Abstract: A simple and rapid method for detecting Plasmodium falciparum in human blood was used in this study. The assay is based on DNA-DNA spot hybridization. For this purpose, the total genomic DNA of P. falciporum was isolated and purified from the parasite cultured in vitro. Then the total genomic DNA was used as a probe and labelled with [α-32P]-dATP by nick-translation. Twenty-five test samples, ten-microliter lysed infected blood each, were spotted onto dry nitrocellulose paper and hybridized with labelled genomic DNA. After hybridization, the paper was exposed to X-ray film for autoradiography, resulting in an image in places where hybridization occurred.The result shows that the assay appears to be sensitive enough to detect parasitaemia up to 0.0009%. No visible hybridization was detected in normal human blood or human leukocytes. Many samples can be processed simultaneously. This may be applicable to mass survey for detecting P. falciparum in epidemiological study (Fig. 2).