Abstract Malaria Rapid Diagnostic Tests (RDTs) plays an important role in malaria management and control. The Pf HRP2 based RDT kit is the most widely used RDT for malaria diagnosis in Nigeria but is affected by the deletion of HRP2 gene in Plasmodium falciparum parasites. Therefore, identifying the prevalence and distribution of Plasmodium falciparum parasites with deleted Pf HRP2 is important for malaria control. Pf HRP2 gene deletion was assessed in this study by first carrying out Giemsa stained thick blood film microscopy and Pf HRP2 RDT strip test. The samples were further analyzed for molecular examination by PCR assay for multiple single–copy genes (Pf Cox3, Pf HRP2, Pf HRP3 and Pf Beta tubulin). This study found the existence of eight (8) Plasmodium falciparum isolates lacking the HRP2 gene in the samples analyzed, this necessitates the need to develop a unique RDT Kit targeting other housekeeping genes unique for Plasmodium falciparum with far greater sensitivity than the current ones as to reduce the chances of false negative RDT result as well as developing unique RDT Kits targeting both PfHRP2 and PfHRP3 genes concomitantly in order to reduce the chances of having a false positive RDT results.