首页> 外文期刊>The American Journal of Tropical Medicine and Hygiene >Short report: Rapid DNA extraction from archive blood spots on filter paper for genotyping of Plasmodium falciparum.
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Short report: Rapid DNA extraction from archive blood spots on filter paper for genotyping of Plasmodium falciparum.

机译:简短报告:从滤纸上存档的血斑中快速提取DNA,用于恶性疟原虫的基因分型。

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摘要

The practical advantages of sampling and storing blood on filter paper for analyses of human and pathogen genes highlight the need for reliable, sensitive, and cost-effective DNA extraction methods. We describe a new Tris-EDTA (TE) buffer-based method for extraction of DNA from blood dried on filter paper. The method was evaluated against the commonly used methanol and Chelex methods, regarding polymerase chain reaction detection of Plasmodium falciparum parasites from samples stored for 1-2 years. The sensitivity of detection was dependent on the parasite density and type of filter paper. For 3MM Whatman filter paper, the sensitivity was 100%, 73%, and 93% for the TE, methanol, and Chelex methods, respectively. For the longer stored 903 Schleicher & Schuell filter paper, the sensitivity was 93%, 73%, and 0%, respectively. This rapid, simple, and inexpensive extraction method generated superior results from archived specimens compared with the two standard methods and may represent a useful tool in molecular epidemiologic studies.
机译:在滤纸上采样和存储血液以分析人和病原体基因的实际优势凸显了对可靠,灵敏和经济高效的DNA提取方法的需求。我们描述了一种新的基于Tris-EDTA(TE)缓冲液的方法,用于从滤纸上干燥的血液中提取DNA。针对存储1-2年的样本中关于恶性疟原虫寄生虫的聚合酶链反应检测,针对常用的甲醇和Chelex方法对该方法进行了评估。检测的灵敏度取决于寄生虫的密度和滤纸的类型。对于3MM Whatman滤纸,TE,甲醇和Chelex方法的灵敏度分别为100%,73%和93%。对于存储时间较长的903 Schleicher&Schuell滤纸,灵敏度分别为93%,73%和0%。与两种标准方法相比,这种快速,简单且廉价的提取方法从存档的标本中产生了优异的结果,并且可能代表了分子流行病学研究中的有用工具。

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