New Diagnostic Approach Based on Loop-Mediated Isothermal Amplification as a Tool for Malaria Epidemiological Surveillance

Relevance. Imported malaria cases are registered annually in Russia, mainly originating from African and Southeast Asian countries. Although there has been no local transmission of the disease within the country since 2018, the risk of importation and spread of the infection remains. This necessitates reliable diagnostic methods for rapid response. Globally, malaria remains one of the most significant public health threats in endemic regions. Since 2007, World Malaria Day has been observed on April 25, underscoring the importance of consolidating efforts in the diagnosis, prevention, and treatment of the disease. Current diagnostic methods, such as microscopy and immunochromatographic tests, have limitations in terms of sensitivity, specificity, and suitability for mass screening.

Aims. To develop and validate a reagent kit for malaria diagnosis based on loop mediated isothermal amplification (LAMP) method, and to assess its diagnostic performance.

Materials and methods. This study presents the development and validation of the AmpliSens® Plasmodium-IT reagent kit for the qualitative detection of DNA from pathogenic malaria Plasmodium species (P. falciparum, P. vivax, P. malariae, P. ovale, P. knowlesi) using LAMP with real-time fluorescent detection. Detection limit was assessed using serial dilutions of Plasmodium DNA. The diagnostic sensitivity and specificity were evaluated using clinical samples of whole venous blood characterized by PCR.

Results. The developed reagent kit demonstrates high analytical sensitivity (detection limit: 103 GE/mL), 100 % diagnostic sensitivity, and 100 % specificity when tested on clinical whole-blood samples. The analysis time is 25–30 minutes, and a positive signal in cases of high parasitemia is detected within 3–12 minutes.

Conclusion. The developed reagent kit is a fast and accurate tool for malaria diagnosis, suitable for use in stationary laboratories for epidemiological surveillance and screening.

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