Mussa Ali, Talib Mustafa, Mohamed Zeehaida, Hajissa Khalid
Genetics and Molecular Biology Laboratory, Department of Zoology, Faculty of Science, University of Khartoum, Khartoum, Sudan.
Department of Zoology, Faculty of Science and Technology, Omdurman Islamic University, B.O.Box382, Omdurman, Sudan.
BMC Res Notes. 2019 Jun 11;12(1):334. doi: 10.1186/s13104-019-4361-6.
Rapid diagnostic tests (RDTs) play a crucial role in the management and control of malaria infection. The histidine-rich protein 2 (PfHRP-2) based RDTs are the most commonly used RDTs for malaria diagnosis in Sudan. Deletion of pfhrp2 in Plasmodium falciparum genome affect the accuracy of PfHRP-2 based RDT kits. This study aimed to identify molecular variation of pfhrp2 among suspected malaria patients from different clinics in Omdurman, Sudan.
A noticeable variation between the RDT (Alltest Biotech, China) and nPCR results was observed, for RDT 78% (46/59) were P. falciparum positive, 6.8% (4/59) were co-infected with both P. falciparum and Plasmodium vivax, 15.3% (9/59) were negative by the RDT. However, when the nPCR was applied only 44.1% (26/59) and 55.9% (33/59) was P. falciparum positive and negative respectively. The pfhrp2 was further amplified form all nPCR positive samples. Only 17 DNA samples were positive from the 26 positive P. falciparum, interestingly, variation in band sizes was observed and further confirmed by DNA sequencing, and sequencing analysis revealed a high-level of genetic diversity of the pfhrp2 gene in the parasite population from the study area. However, despite extreme sequence variation, diversity of PfHRP2 does not appear to affect RDT performance.
快速诊断检测(RDTs)在疟疾感染的管理和控制中发挥着关键作用。基于富含组氨酸蛋白2(PfHRP - 2)的RDTs是苏丹最常用的疟疾诊断RDTs。恶性疟原虫基因组中pfhrp2的缺失会影响基于PfHRP - 2的RDT试剂盒的准确性。本研究旨在确定苏丹恩图曼不同诊所疑似疟疾患者中pfhrp2的分子变异情况。
观察到RDT(中国奥泰生物科技公司)与巢式聚合酶链反应(nPCR)结果之间存在显著差异,RDT检测中78%(46/59)为恶性疟原虫阳性,6.8%(4/59)同时感染恶性疟原虫和间日疟原虫,15.3%(9/59)RDT检测为阴性。然而,当应用nPCR时,分别有44.1%(26/59)为恶性疟原虫阳性,55.9%(33/59)为阴性。从所有nPCR阳性样本中进一步扩增pfhrp2。在26例阳性恶性疟原虫样本中只有17个DNA样本呈阳性,有趣的是,观察到条带大小存在差异,并通过DNA测序进一步证实,测序分析显示研究区域寄生虫群体中pfhrp2基因具有高度的遗传多样性。然而,尽管序列变异极大,但PfHRP2的多样性似乎并未影响RDT的性能。