Mitogen-activated protein kinase 5, a novel molecular marker for the identification and detection of Trypanozoon...

Based on the sequence of mitogen-activated protein kinase 5 (TbMAPK5) gene, we have developed a specific PCR method which can delineate the species within the Trypanozoon subgenus from other parasites or host DNA conveniently. In view of further application in field studies, we performed loop-mediated isothermal amplification (LAMP) employing filter paper flecked with rodent blood. Our data showed that TbMAPK5-specific LAMP was sensitive enough to detect a very low parasitemia during the early stages of the infection and fluctuating parasitemia period, and is below the detection limit by microscope. The detection limit for the infected blood sample was 1000 trypanosomes/ml of blood. Based on these results, we consider that TbMAPK5 locus may be a useful target for LAMP diagnosis providing sensitivity and the potential for the genotyping/identification of Trypanozoon species.