Evaluation of Visual LAMP detection of Plasmodium falciparum and non-Plasmodium falciparum

Abstract

Background: To detect the DNA of the malaria using Loop-mediated isothermal amplification (LAMP) and evaluate the effect. Method; According to the 18s rRNA sequence of the malaria, the LAMP primer of Plasmodium falciparum and non-Plasmodium falciparum were designed using Primer Explorer V5 software. The method of Visual LAMP detecting malaria was formation. The sensitivity, specificity and amplification efficiency were tested, compared with the Nest-PCR. Result: The filter papers of blood from 47 patients with Plasmodium falciparum and 49 with non-Plasmodium falciparum were detected using Visual LAMP. The patients with Plasmodium falciparum were all positive. In the patients with non-Plasmodium falciparum , false negative rate was 2.1%. In the 10 patients with Leishmaniasis, 8 with Schistosomiasis japonicum, 38 healthy human, none was positive. The results using visual LAMP were consistent with Nest-PCR (Kappa value = 0.956). All of the Tt in Visual LAMP are less than 60 min, especially the Tt of Plasmodium falciparum is 18.2 min. When adding calcium pyridoxine indicator during the Visual LAMP detection, the progress of the reaction to different substes of malaria delayed of about 9-23 min. Conclusion: LAMP is efficient and visible, deserved the prospect of application in the on-site and primary medical institutions.