Genetic Diversity of Plasmodium falciparum and Plasmodium vivax Field Isolates from the Nowshera District of Pakistan

Background The genetic diversity of malaria parasites contributes to their ability to adapt to environmental changes, develop drug resistance and escape from the host immune system; hence, it is very important for control measures of malaria. This study aimed to analyse the genetic diversity of the pfmsp1 and pfmsp2 genes in P. falciparum and the Pvmsp-3α gene in P. vivax isolates from District Nowshera in Pakistan. Methods Blood samples from 124 consenting patients with uncomplicated malaria presenting to different hospitals of district Nowshera were collected during March-August 2019, representing 28 P. falciparum and 96 P. vivax isolates. DNA from all samples was subjected to nested PCR-based allele-specific marker analysis. Pvmsp-3α amplified fragments were further treated with restriction fragment length polymorphism (RFLP)-based Hha1 restriction enzyme. Results In P. falciparum , 21 alleles were detected, including 14 alleles for Pfmsp-1 and 7 alleles for Pfmsp-2 . The suballelic families MAD20 (50%) in the Pfmsp-1 family and FC27 (75%) in the Pfmsp-2 family were predominant. The multiplicity of infection (MOI) was calculated as 1.4 and 1.2 for Pfmsp-1 and Pfmsp - 2 , respectively, with an overall mean MOI of 1.34. In P. vivax , 4 allelic variants, Type A-D, were detected for Pvmsp- 3α through nested PCR, while after RFLP digestion of amplicons, 9 suballelic variants (A1-A4, B1, B2, C1, C2 and D1) were observed at the Pvmsp-3α locus. Conclusion This first ever report of molecular characterization of P. falciparum and P. vivax genotypes from District Nowshera, Pakistan reveals moderate to high allelic diversity in these parasites from District Nowshera, Pakistan.