Loop-mediated isothermal amplification for detection of African trypanosomes
Loop-mediated isothermal amplification for detection of African trypanosomes
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DOI：
10.1128/jcm.41.12.5517-5524.2003
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发表时间：
2003-12-01
影响因子：
9.4
通讯作者：
Igarashi, I
中科院分区：
文献类型：
--
作者：
Kuboki, N;Inoue, N;Igarashi, I
虽然聚合酶链式反应是在人和动物中检测非洲锥虫的一种选择方法，但这种方法的费用使其无法用作检测非洲国家地方性锥虫病的诊断方法。环介导的等温扩增(LAMP)反应是一种仅用简单的孵化器就能在等温条件下高特异性、高效、快速地扩增DNA的方法。与基于聚合酶链式反应的方法相比，LAMP的另一个优势是可以用分光光度和/或肉眼监测DNA扩增，而无需使用染料。在这里，我们报告了一种基于LAMP技术的高灵敏度、特异性和简单易行的诊断方法，用于检测布氏锥虫种群(包括布氏锥虫、冈比亚布氏锥虫、罗氏布氏锥虫和伊氏锥虫)和孔氏锥虫的寄生虫。我们发现，基于LAMP的方法在体外检测锥虫的灵敏度比基于PCR的方法高100倍。对感染人类感染的布氏冈比亚毛滴虫的小鼠进行的体内研究进一步强调了LAMP作为非洲锥虫病诊断工具的潜在临床重要性。
While PCR is a method of choice for the detection of African trypanosomes in both humans and animals, the expense of this method negates its use as a diagnostic method for the detection of endemic trypanosomiasis in African countries. The loop-mediated isothermal amplification (LAMP) reaction is a method that amplifies DNA with high specificity, efficiency, and rapidity under isothermal conditions with only simple incubators. An added advantage of LAMP over PCR-based methods is that DNA amplification can be monitored spectrophotometrically and/or with the naked eye without the use of dyes. Here we report our conditions for a highly sensitive, specific, and easy diagnostic assay based on LAMP technology for the detection of parasites in the Trypanosoma brucei group (including T. brucei brucei, T. brucei gambiense, T. brucei rhodesiense, and T. evansi) and T. congolense. We show that the sensitivity of the LAMP-based method for detection of trypanosomes in vitro is up to 100 times higher than that of PCR-based methods. In vivo studies in mice infected with human-infective T. brucei gambiense further highlight the potential clinical importance of LAMP as a diagnostic tool for the identification of African trypanosomiasis.