Department of Molecular Biology, Virtual University of Pakistan, Lahore, Pakistan.
Department of Medicine, Polyclinic Hospital, Islamabad, Pakistan.
Malar J. 2021 Feb 25;20(1):112. doi: 10.1186/s12936-021-03654-w.
Plasmodium vivax contributes to over 70% malaria burden in Pakistan, but limited data exists on various aspects including genetic diversity of the parasite as compared to other parts of the world. Since the information about the genetic diversity of P. vivax assists to understand the population dynamics of the parasite, the current study was designed to understand population divergence of P. vivax in Pakistan using circumsporozoite protein (pvcsp) and merozoite surface protein-1 (pvmsp-1) genes as molecular markers.
The PCR for pvcsp and pvmsp-1 genes was carried out for 150 P. vivax isolates, followed by DNA sequencing of 35 and 30, respectively. Genetic diversity and polymorphism were analysed using ChromasPro, ClustalW, MEGA7, DnaSP v.5 and WebLogo programs.
The PCR for pvcsp and pvmsp-1 genes was carried out for 150 P. vivax isolates and resulting the PCR products of 1100 bp for pvcsp and ~ 400 bp for pvmsp-1 genes, respectively. In the central-repeat region (CRR) of pvcsp gene, sequences comprised of four variable repeats of PRMs, out of which GDRADGQPA (PRM1), GDRAAGQPA (PRM2) were more extensively dispersed among the P. vivax isolates. Partial sequences (~ 400 bp) of block 2 of pvmsp-1 gene depicted high level of diversity.
The results revealed the polymorphism and genetic diversity especially at the CRR of pvcsp and block 2 of pvmsp-1 genes, respectively. The base-line data presented here warrants future studies to investigate more into the genetic diversity of P. vivax with large sample size from across the country for better understanding of population dynamics of P. vivax that will help to control malaria at individual and community level.
间日疟原虫在巴基斯坦造成超过 70%的疟疾负担,但与世界其他地区相比,有关其寄生虫的各种方面的信息有限,包括遗传多样性。由于了解间日疟原虫的遗传多样性有助于了解寄生虫的种群动态,因此本研究旨在使用环子孢子蛋白(pvcsp)和裂殖子表面蛋白-1(pvmsp-1)基因作为分子标记来了解巴基斯坦间日疟原虫的种群分化。
对 150 株间日疟原虫分离株进行 pvcsp 和 pvmsp-1 基因的 PCR 扩增,随后分别对 35 株和 30 株进行 DNA 测序。使用 ChromasPro、ClustalW、MEGA7、DnaSP v.5 和 WebLogo 程序分析遗传多样性和多态性。
对 150 株间日疟原虫分离株进行 pvcsp 和 pvmsp-1 基因的 PCR 扩增,得到 pvcsp 基因的 1100bp 产物和 pvmsp-1 基因的400bp 产物。在 pvcsp 基因的中央重复区(CRR)中,序列由 PRMs 的四个可变重复组成,其中 GDRADGQPA(PRM1)和 GDRAAGQPA(PRM2)在间日疟原虫分离株中分布更为广泛。pvmsp-1 基因的第 2 块的部分序列(400bp)显示出高度的多样性。
结果显示,pvcsp 的 CRR 和 pvmsp-1 的第 2 块基因分别具有遗传多态性和遗传多样性。本研究提供了基础数据,未来需要进行更多研究,以了解全国范围内更大样本量的间日疟原虫的遗传多样性,从而更好地了解间日疟原虫的种群动态,有助于在个人和社区层面控制疟疾。