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恶性疟原虫在干血斑上的聚合酶链反应扩增

Polymerase chain reaction amplification from Plasmodium falciparum on dried blood spots.

作者信息

Long G W, Fries L, Watt G H, Hoffman S L

机构信息

Center for Vaccine Research, School of Hygiene and Public Health, Johns Hopkins University, Baltimore, Maryland, USA.

出版信息

Am J Trop Med Hyg. 1995 Apr;52(4):344-6. doi: 10.4269/ajtmh.1995.52.344.

Abstract

We report a simple method for the polymerase chain reaction (PCR) amplification of whole blood samples collected on filter paper. The blood spot was used directly in the PCR after treatment with methanol. We evaluated this assay using clinical samples collected from subjects in a Plasmodium falciparum vaccine trial and from samples collected during a hospital-based study in Thailand. Specimens prepared from heparinized blood samples were successfully amplified following pretreatment with heparinase. Sensitivity was 100% when compared with thick blood film results in the vaccine trial (range = 4-60 parasites/microliters, median = 8/microliters) and 94.6% (range = 3-133,988 parasites/microliters, median = 616/microliters) in the hospital study.

摘要

我们报告了一种用于对滤纸上采集的全血样本进行聚合酶链反应(PCR)扩增的简单方法。血斑经甲醇处理后直接用于PCR。我们使用从恶性疟原虫疫苗试验受试者采集的临床样本以及在泰国一项基于医院的研究中采集的样本对该检测方法进行了评估。用肝素酶预处理后,从肝素化血样制备的标本成功实现了扩增。与疫苗试验中的厚血膜结果相比,灵敏度为100%(范围 = 4 - 60个寄生虫/微升,中位数 = 8/微升),在医院研究中为94.6%(范围 = 3 - 133,988个寄生虫/微升,中位数 = 616/微升)。

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