中国寄生虫学与寄生虫病杂志 ›› 1995, Vol. 13 ›› Issue (2): 120-126.

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滤纸干血滴抽提恶性疟原虫 DNA 用于 PCR 扩增

张龙兴,詹斌,王聚君,冯晓平   

  1. 中国预防医学科学院寄生虫病研究所
  • 出版日期:1995-05-31 发布日期:2017-01-05

APPLICATION OF PLASMODIUM FALCIPARUM DNA EXTRACT FROM DRIED BLOOD SPOT OF FALCIPARUM MALARIA PATIENT IN POLYMERASE CHAIN REACTION

  • Online:1995-05-31 Published:2017-01-05

摘要: 以STE-蛋白酶K-SDS、甲醇-蛋白酶K-SDS、Chelex-100加热法和Chelex-100蛋白酶K等四种方法对恶性疟患者滤纸干血滴样品进行DNA抽提,供PCR扩增特异性AMA-1DNA片段。结果显示,除STE-蛋白酶K-SDS法抽提DNA进行PCR试验未能获得扩增产物外,其他3种方法抽提DNA后进行PCR试验均获得特异性约900bpDNA片段,可供进一步试验。

关键词: 恶性疟原虫, 滤纸干血滴, 蛋白酶, 抽提方法, 凝胶电泳, 金属离子, 琼脂糖, 弱阳离子交换剂, 采血方法, 特异性

Abstract: Polymerase chain reaction (PCR) primered with primers 3,4 and 5,6 was performed using DNA extracted from dried blood spot of a falciparum malaria patient from Mengpeng Township,Yunnan using four different methods,including STE proteinase K SDS,methanol proteinase K SDS,Chelex 100 and Chelex 100 proteinase K to amplify DNA of apical membrane antigen 1(AMA 1).A 900 bp DNA band could be seen on the ethidium bromide stained agarose gel electrophoresis of the PCR products when DNA was extracted using all the above mentioned methods except STE proteinase K SDS method. DNA extracted with Chelax 100 was recommended.

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