Molecular Investigation of FIKK Kinase Family to Develop PCR-Based Diagnosis of Plasmodium falciparum.

Accurate malaria diagnosis is crucial for effective disease management as different Plasmodium species require specific treatment regimens. Current detection methods have limitations related to sensitivity and specificity. This is mainly due to employing similar targets such as 18S rRNA, Pf-ldh, Pf-hrp-2, and aldolase with significant homology to human counterparts. Targeting Plasmodium fikk kinases that are unique to P. falciparum offers a novel approach for developing potential biomarkers. We have identified exclusive regions of fikk kinases using in-silico PCR and later validated our findings using in-vitro PCR. We observed exceptional sensitivity with our designed primers of the targeted fikk kinases, with the detection limit going as low as 10^-5 ng level of parasite DNA and 0.0003% parasitemia. The shortlisted primers also selectively identified P. falciparum in the presence of Plasmodium vivax or several other bacterial, viral, and fungal pathogens. Detection of mock patient samples indicates that the fikk-based PCR diagnosis is giving accurate results, and it is much better than the existing method. Thus, we show that the fikk kinases from P. falciparum are excellent targets for developing novel biomarkers with high sensitivity and specificity.