Simple, Field-Usable Fluorescence-Based Isothermal LAMP Assay for the On-Site Diagnosis of Malaria

CDC researchers have developed improved Loop-Mediated Isothermal Amplification (LAMP) assays for the nucleic acid-based diagnosis of malaria in field settings. The approach employs Plasmodium genus-specific LAMP primers and a portable tube scanner to run the LAMP reaction and measure fluorescence signal (e.g., SYBR green) as a measure of DNA amplification in real time. Using this platform, the researchers were able to detect several different species of the human malaria parasites.

This LAMP approach has demonstrated the ability of this assay to detect P. falciparum in clinical samples with approximately 99-100% sensitivity, an improvement over standard nested PCR methodology. Additionally, DNA extracted by direct boiling of whole blood works well in this assay. Further, several important practical aspects of the method make it an attractive method for on-site uses. For example, it is portable to remote locales, required equipment is easily powered by battery or other alternate power sources, and no post-assay tasks, such as gel electrophoresis, are required. This method is also technically easier to perform than nested PCR, has automation features allowing direct reporting of results from remote locations, and can be modified to handle large sample numbers.