中国寄生虫学与寄生虫病杂志 ›› 1994, Vol. 12 ›› Issue (3): 169-171.

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恶性疟原虫SSUrDNA特定片段的体外扩增及鉴定

万磊,陈培霞,薛采芳,刘忠湘,姜绍谆   

  1. 第四军医大学寄生虫学教研室; 第四军医大学微生物学教研室
  • 出版日期:1994-08-31 发布日期:2017-01-06
  • 基金资助:
    国家自然科学基金

AMPLIFICATION IN VITRO AND IDENTIFICATION OF SMALL SUBUNIT RIBOSOMAL DNA FRAGMENT OF PLASMODIUM FALCIPARUM 

  • Online:1994-08-31 Published:2017-01-06

摘要: 根据已知疟原虫、其它寄生人体原虫及人的小亚单位核糖体核糖核酸基因(SSUrDNA)编码区序列,借助计算机核酸序列软件分析,设计出一对用于恶性疟原虫(P.f.)SSUrDNA特定片段扩增的引物。采用双温度点聚合酶链反应(PCR)技术,从我国P.f.FCC/YN茅芽株红内期基因组DNA中,扩增出约570个碱基对(bp)的DNA片段,与预期长度相符;而间日疟原虫(P.v.)、利什曼原虫(L.d.)、弓形虫(T.g.)及人白细胞DNA样本均无此扩增带出现。扩增片段经限制性内切酶消化和Northern印迹杂交分析,证实为P.f.SSUrDNA目的片段。

关键词: 恶性疟原虫, 小亚单位核糖体核糖核酸基因, 聚合酶链反应, 限制性内切酶消化, Northern印迹杂交

Abstract: According to computer analysis of SSUrDNA sequences of Plasmodium ,other protozooa and human,two oligonucleotide primers were designed.A DNA fragment,about 570 base pairs,was successfully amplified by two temperature point polymerase chain reaction from the genomic DNA of cultivated erythrocytic stage of P.falciparum FCC/YN(Simao),but no fragment was obtained from that of P.vivax,L.donovani,T.gondii and humans.It has been confirmed that the amplified fragment was indeed expected SSUrDNA segment of P.falciparum by means of restriction endonuclease digestions and Northern blot hybridization.

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