AUTHOR=Khan Shahid Niaz , Ali Rehman , Khan Sanaullah , Rooman Muhammad , Norin Sadia , Zareen Shehzad , Ali Ijaz , Ayaz Sultan 

TITLE=Genetic Diversity of Polymorphic Marker Merozoite Surface Protein 1 (Msp-1) and 2 (Msp-2) Genes of Plasmodium falciparum Isolates From Malaria Endemic Region of Pakistan

JOURNAL=Frontiers in Genetics

VOLUME=12

YEAR=2021

URL=https://www.frontiersin.org/journals/genetics/articles/10.3389/fgene.2021.751552

DOI=10.3389/fgene.2021.751552

ISSN=1664-8021

ABSTRACT=<p><bold>Background:</bold> Understanding the genetic diversity of <italic>Plasmodium</italic> species through polymorphic studies can assist in designing more effective control strategies of malaria like new drug formulation and development of a vaccine. Pakistan is moderate endemic for <italic>Plasmodium falciparum</italic>, but little is known about the genetic diversity of this parasite. This study aimed to investigate the molecular diversity of <italic>P. falciparum</italic> based on <italic>msp-1</italic> and <italic>msp-2</italic> genes in the malaria-endemic regions of Khyber Pakhtunkhwa, Pakistan.</p><p><bold>Methods:</bold> A total of 199/723 blood samples, tested positive by microscopy for <italic>falciparum</italic> malaria, were collected from four districts (Dera Ismail Khan, Karak, Mardan, and Peshawar) of Khyber Pakhtunkhwa. Nested PCR amplification technique was employed to target block 2 of <italic>msp-1</italic> and the central domain of <italic>msp-2</italic> genes, including their respective allelic families K1, MAD20, RO33, FC27, and 3D7/IC, and to detect the extent of genetic diversity of <italic>P. falciparum</italic> clinical isolates.</p><p><bold>Results:</bold> Among the 199 microscopy-positive <italic>P. falciparum</italic> samples, a total of 192 were confirmed using PCR. Ninety-seven amplicons were observed for <italic>msp</italic>-<italic>1</italic> and 95 for <italic>msp-2</italic>. A total of 33 genotypes, 17 for <italic>msp-1</italic> (eight K1, six MAD20, and three RO33) and 16 for <italic>msp-2</italic> (nine FC27 and seven 3D7/IC), were identified. The specific allelic frequency of the K1 family was higher (44.3%) than that of MAD20 (33.0%) and RO33 (23.0%) for <italic>msp-1</italic>, while the FC27 allelic family was dominant (60.0%) compared with 3D7/IC (40.0%) for <italic>msp-2</italic>. No polyclonal infection was observed in <italic>msp-1</italic> and <italic>msp-2</italic>. The expected heterozygosity was 0.98 and 0.97 for <italic>msp-1</italic> and <italic>msp-2</italic>, respectively.</p><p><bold>Conclusion:</bold> It was concluded that the <italic>P. falciparum</italic> populations are highly polymorphic, and diverse allelic variants of <italic>msp-1</italic> and <italic>msp-2</italic> are present in Khyber Pakhtunkhwa, Pakistan.</p>