Loop-mediated isothermal PCR (LAMP) for the diagnosis of falciparum malaria
Loop-mediated isothermal PCR (LAMP) for the diagnosis of falciparum malaria
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DOI：
10.4269/ajtmh.2007.77.972
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发表时间：
2007-11-01
影响因子：
3.3
通讯作者：
Dondorp, Arjen M.
中科院分区：
文献类型：
--
作者：
Paris, Daniel H.;Imwong, Mallika;Dondorp, Arjen M.
最近描述的用于恶性疟原虫分子检测的环介导等温聚合酶链反应 (LAMP) 与显微镜、基于 PfHRP2 的快速诊断测试 (RDT) 和巢式聚合酶链反应 (PCR) 进行了比较，作为 115 名孟加拉国发烧住院患者的“金标准”。 LAMP的DNA提取通过常规方法或简单加热样品进行；测试结果通过目视或凝胶电泳评估。常规 DNA 提取和凝胶电泳与参考方法的一致性最高 (81.7%，kappa = 0.64)，灵敏度 (95% CI) 为 76.1% (68.3-83.9%)，与 RDT 和显微镜检查相当，但特异性为 89.6% (84.0-95.2%)，而 RDT 和显微镜检查的特异性为 100%。通过热处理提取 DNA 会使特异性降低到不可接受的水平。 LAMP 能够在技术资源有限的情况下对恶性疟疾进行分子诊断，但需要进一步优化。该结果与早期研究报告的较高准确度形成鲜明对比，该研究将 LAMP 与未经验证的 PCR 方法进行比较。
A recently described loop-mediated isothermal polymerase chain reaction (LAMP) for molecular detection of Plasmodium falciparum was compared with microscopy, PfHRP2-based rapid diagnostic test (RDT), and nested polymerase chain reaction (PCR) as the "gold standard" in 115 Bangladeshi in-patients with fever. DNA extraction for LAMP was conducted by conventional methods or simple heating of the sample; test results were either assessed visually or by gel electrophoresis. Conventional DNA extraction followed by gel electrophoresis had the highest agreement with the reference method (81.7%, kappa = 0.64), with a sensitivity (95% CI) of 76.1% (68.3-83.9%), comparable to RDT and microscopy, but a specificity of 89.6% (84.0-95.2%) compared with 100% for RDT and microscopy. DNA extraction by heat treatment deteriorated specificity to unacceptable levels. LAMP enables molecular diagnosis of falciparum malaria in settings with limited technical resources but will need further optimization. The results are in contrast with a higher accuracy reported in an earlier study comparing LAMP with a non-validated PCR method.