Detection of Mycobacterium ulcerans by the Loop Mediated Isothermal Amplification Method

In order to develop a simple and rapid test that can be used to diagnose Buruli ulcer under field conditions, we modified the conventional LAMP assay by using a disposable pocket warmer as a heating device for generating a constant temperature for the test reaction and employed the use of crude sample preparations consisting of boiled and unboiled extracts of the clinical specimen instead of using purified DNA as the diagnostic specimen. Thirty clinical specimens from suspected Buruli ulcer patients were investigated by the modified LAMP (or pocket warmer LAMP) and the conventional LAMP, as well as IS2404 PCR, a reference method for the detection of Mycobacterium ulcerans. There was no significant difference in the detection rate (63–70%) in all of the methods when purified samples were used for the tests. On the other hand the use of crude specimen preparation resulted in a drop in detection rate (30–40%). This study demonstrates that the LAMP test can be used for rapid detection of M. ulcerans when purified DNA preparations are used. With further improvements in the sample reaction, as well as in specimen purification, the pocket warmer LAMP may provide a simple and rapid diagnostic test for Buruli ulcer

Comparison of IS<i>2404</i> PCR with pocket warmer LAMP for the detection of <i>Mycobacterium ulcerans</i> using different DNA templates.

*<p>Sensitivity as compared with IS<i>2404</i> PCR.</p><p>LAMP (Loop mediated isothermal amplification).</p

Comparison of IS<i>2404</i> PCR with conventional LAMP for the detection of <i>Mycobacterium ulcerans</i> using different DNA templates.

*<p>Sensitivity as compared with IS<i>2404</i> PCR</p><p>LAMP (Loop mediated isothermal amplification).</p