Khan Shahid Niaz, Ali Rehman, Khan Sanaullah, Rooman Muhammad, Norin Sadia, Zareen Shehzad, Ali Ijaz, Ayaz Sultan
Department of Zoology, Faculty of Biological Sciences, Kohat University of Science and Technology, Kohat, Pakistan.
Department of Zoology, University of Peshawar, Peshawar, Pakistan.
Front Genet. 2021 Nov 17;12:751552. doi: 10.3389/fgene.2021.751552. eCollection 2021.
Understanding the genetic diversity of 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 , but little is known about the genetic diversity of this parasite. This study aimed to investigate the molecular diversity of based on and genes in the malaria-endemic regions of Khyber Pakhtunkhwa, Pakistan. A total of 199/723 blood samples, tested positive by microscopy for 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 and the central domain of genes, including their respective allelic families K1, MAD20, RO33, FC27, and 3D7/IC, and to detect the extent of genetic diversity of clinical isolates. Among the 199 microscopy-positive samples, a total of 192 were confirmed using PCR. Ninety-seven amplicons were observed for - and 95 for . A total of 33 genotypes, 17 for (eight K1, six MAD20, and three RO33) and 16 for (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 , while the FC27 allelic family was dominant (60.0%) compared with 3D7/IC (40.0%) for . No polyclonal infection was observed in and . The expected heterozygosity was 0.98 and 0.97 for and , respectively. It was concluded that the populations are highly polymorphic, and diverse allelic variants of and are present in Khyber Pakhtunkhwa, Pakistan.
通过多态性研究了解物种的遗传多样性有助于设计更有效的疟疾控制策略，如新药配方和疫苗研发。巴基斯坦是疟疾中度流行地区，但对这种寄生虫的遗传多样性知之甚少。本研究旨在基于巴基斯坦开伯尔-普赫图赫瓦省疟疾流行地区的msp1和msp2基因，调查疟原虫的分子多样性。从开伯尔-普赫图赫瓦省的四个地区（德拉伊斯梅尔汗、卡拉奇、马尔丹和白沙瓦）共采集了199/723份经显微镜检测为疟原虫阳性的血样。采用巢式PCR扩增技术靶向msp1的第2区段和msp2基因的中央结构域，包括它们各自的等位基因家族K1、MAD20、RO33、FC27和3D7/IC，并检测疟原虫临床分离株的遗传多样性程度。在199份显微镜检测阳性的疟原虫样本中，共有192份通过PCR得到确认。msp1观察到97个扩增子，msp2观察到95个扩增子。共鉴定出33种基因型，msp1为17种（8种K1、6种MAD20和3种RO33），msp2为16种（9种FC27和7种3D7/IC）。msp1中K1家族的特定等位基因频率（44.3%）高于MAD20（33.0%）和RO33（23.0%），而msp2中FC27等位基因家族（60.0%）比3D7/IC（40.0%）更占优势。在msp1和msp2中均未观察到多克隆感染。msp1和msp2的预期杂合度分别为0.98和0.97。得出的结论是，疟原虫群体具有高度多态性，巴基斯坦开伯尔-普赫图赫瓦省存在msp1和msp2的多种等位基因变体。
