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Background  Loop-mediated isothermal amplification (LAMP) for malaria diagnosis at the point of care (POC) depends on the detection capacity of synthesized nucleic acids and the specificity of the amplification target. To improve malaria diagnosis, new colorimetric LAMP tests were developed using multicopy targets for  Plasmodium vivax  and  Plasmodium falciparum  detection.    Methods  The cytochrome oxidase I ( COX1 ) mitochondrial gene and the non-coding sequence  Pvr47  for  P. vivax , and the sub-telomeric sequence of erythrocyte membrane protein 1 ( EMP1 ) and the non-coding sequence  Pfr364  for  P. falciparum  were targeted to design new LAMP primers. The limit of detection (LOD) of each colorimetric LAMP was established and assessed with DNA extracted by mini spin column kit and the Boil & Spin method from 28 microscopy infections, 101 malaria submicroscopic infections detected by real-time PCR only, and 183 negatives infections by both microscopy and PCR.    Results  The LODs for the colorimetric LAMPs were estimated between 2.4 to 3.7 parasites/µL of whole blood. For  P. vivax  detection, the colorimetric LAMP using the COX1 target showed a better performance than the  Pvr47  target, whereas the  Pfr364  target was the most specific for  P. falciparum  detection. All microscopic infections of  P. vivax  were detected by PvCOX1-LAMP using the mini spin column kit DNA extraction method and 81% (17/21) were detected using Boil & Spin sample preparation. Moreover, all microscopic infections of  P. falciparum  were detected by Pfr364-LAMP using both sample preparation methods. In total, PvCOX1-LAMP and Pfr364-LAMP detected 80.2% (81 samples) of the submicroscopic infections using the DNA extraction method by mini spin column kit, while 36.6% (37 samples) were detected using the Boil & Spin sample preparation method.    Conclusion  The colorimetric LAMPs with multicopy targets using the  COX1  target for  P. vivax  and the  Pfr364  for  P. falciparum  have a high potential to improve POC malaria diagnosis detecting a greater number of submicroscopic  Plasmodium  infections.    Supplementary Information  The online version contains supplementary material available at 10.1186/s12936-021-03753-8.

Multicopy targets for Plasmodium vivax and Plasmodium falciparum detection by colorimetric LAMP