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Sensitive RT-LAMP detection of malaria infection using GelGreen® dye

There is a critical need for sensitive and field deployable diagnostic tools that can detect submicroscopic, but yet transmissible levels of malaria parasites. In a recent issue of PLoS ONE, collaborative team from the Centre Pasteur du Cameroun (Cameroon), the University of Douala (Cameroon), and PATH (Seattle, USA), developed a rapid, sensitive and specific method for Plasmodium falciparum malaria diagnosis based on reverse transcription loop-mediated isothermal amplification (RT-LAMP).

This assay could detect P. falciparum infection in as little as 0.25 ng of total parasite RNA, and had a detection limit of 0.08 parasites/μL (microscopically confirmed) when testing infected whole blood lysates. In most cases, the RT-LAMP reaction was complete within twenty minutes, with some samples showing P. falciparum positivity eight minutes into the assay. To demonstrate the feasibility of the isothermal amplification technique in the field, in a resource-limited setting, the RT-LAMP reactions were successfully performed using an inexpensive and electricity-free system based on the exothermic reaction between common salt and Mg-Fe alloy.

The authors showed the effectiveness of GelGreen®  dye for detection of the RT-LAMP reaction, by either adding the GelGreen® Nucleic Acid Gel Stain to the RT-LAMP reaction tubes before or after amplification, and by viewing the resulting fluorescence by hand-held UV flashlight or laboratory UV light box.

To read the original article, click here.

Kemleu S, Guelig D, Eboumbou Moukoko C, Essangui E, Diesburg S, Mouliom A, et al. A Field-Tailored Reverse Transcription Loop-Mediated Isothermal Assay for High Sensitivity Detection of Plasmodium falciparum Infections. 2016, PLOS ONE 11(11), Nov 8;11(11):e0165506. doi: 10.1371/journal.pone.0165506.

To learn more about the safe and sensitive DNA gel stains GelGreen® and GelRed®, click here.