Lateral Flow Recombinase Polymerase Amplification Assays for the Detection of Human Plasmodium Species

This study highlights the development of two lateral flow recombinase polymerase amplification assays for the diagnosis of human malaria. The lateral flow cassettes contained test lines that captured biotin-, 6-carboxyfluorescein, digoxigenin-, cyanine 5-, and dinitrophenyl-labeled amplicons. The overall process can be completed in 30 minutes. Recombinase polymerase amplification coupled with lateral flow had a detection limit of 1 copy/mL for Plasmodium knowlesi, Plasmodium vivax, and Plasmodium falciparum. No cross-reactivity was observed among nonhuman malaria parasites such as Plasmodium coatneyi, Plasmodium cynomolgi, Plasmodium brasilanium, Plasmodium inui, Plasmodium fragile, Toxoplasma gondii, Sarcocystis spp., Brugia spp., and 20 healthy donors. It is rapid, highly sensitive, robust, and easy to use. The result can be read without the need for special equipment and thus has the potential to serve as an effective alternative to polymerase chain reaction methods for the diagnosis of malaria.

Automated summary

This study developed two lateral flow recombinase polymerase amplification assays for detecting human malaria. The assays showed high sensitivity, with a detection limit of 1 copy/mL for various malaria species. They also demonstrated no cross-reactivity with nonhuman malaria parasites and were easy to use without special equipment. These assays have the potential to be an effective alternative to polymerase chain reaction methods for malaria diagnosis.