Mitochondrial DNA Targets Increase Sensitivity of Malaria Detection Using Loop-Mediated Isothermal Amplification
Mitochondrial DNA Targets Increase Sensitivity of Malaria Detection Using Loop-Mediated Isothermal Amplification
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DOI：
10.1128/jcm.00355-10
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发表时间：
2010-08-01
影响因子：
9.4
通讯作者：
Sutherland, Colin J.
中科院分区：
文献类型：
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作者：
Polley, Spencer D.;Mori, Yasuyoshi;Sutherland, Colin J.
环介导的DNA等温扩增(LAMP)提供了在最小的组织样本处理后检测非常少量的病原体DNA的能力，因此是一种有吸引力的护理点诊断方法。以前利用血液样本和LAMP诊断疟疾的尝试都是针对寄生虫小亚单位rRNA基因，因此对恶性疟原虫的敏感性为每亩约100只寄生虫L。在这里，我们描述了基于LAMP的线粒体靶标用于检测任何疟原虫属寄生虫和特定的恶性疟原虫。这些新的目标允许对每亩L血液中含有5种寄生虫的样本进行常规扩增。扩增在30到40分钟内完成，并通过实时比浊法进行评估，从而提供比最熟练的显微镜专家或使用侧向流动免疫分析进行抗原检测更灵敏的快速诊断。
Loop-mediated isothermal amplification (LAMP) of DNA offers the ability to detect very small quantities of pathogen DNA following minimal tissue sample processing and is thus an attractive methodology for point-of- care diagnostics. Previous attempts to diagnose malaria by the use of blood samples and LAMP have targeted the parasite small-subunit rRNA gene, with a resultant sensitivity for Plasmodium falciparum of around 100 parasites per mu l. Here we describe the use of mitochondrial targets for LAMP-based detection of any Plasmodium genus parasite and of P. falciparum specifically. These new targets allow routine amplification from samples containing as few as five parasites per mu l of blood. Amplification is complete within 30 to 40 min and is assessed by real-time turbidimetry, thereby offering rapid diagnosis with greater sensitivity than is achieved by the most skilled microscopist or antigen detection using lateral flow immunoassays.