Molecular detection and quantification of Plasmodium vivax DNA in blood pellet and plasma samples from patients in Senegal

Babacar Souleymane Sambe, A. Diagne, Hélène Ataume Mawounge Diatta, F. M. Gaba, I. Sarr, Arona Sabène Diatta, S. Diaw, Rokhaya Sané, B. Diouf, I. Vigan-Womas, B. Mbengue, Makhtar Niang
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Abstract

Background The first discovery of Plasmodium vivax infections in Senegal used archived patients’ sera in place of blood pellet, the preferred specimen for the molecular diagnosis of Plasmodium species. The present study assessed the reliability of detecting P. vivax DNA in plasma in comparison to blood pellet from the same patient’s samples. Methods A total of 616 blood samples obtained from febrile patients living in Kolda (2015 and 2020), Tambacounda (2017 and 2020), and Kedougou (2020) regions in Senegal, were first screened for Plasmodium species composition by 18S ssrRNA-based nested PCR. Paired blood pellets and plasma samples were selected from a subset of 50 P. vivax-positive patients matched by age and sex with 50 P. vivax-negative patients, and subjected to a cytochrome b-based qPCR to compare the detection and quantification of P. vivax genomic DNA between the two specimen types. Results and discussion The study reports 1.8% and 14.77% of single and mixed P. vivax infections in the study population, and a high concordance (84%) between the qPCR detection of P. vivax genomic DNA from paired blood pellets and plasma samples. Importantly, all P. vivax negative samples from the blood pellets were also confirmed plasma-negative, and parasitaemia in blood pellets was higher compared to plasma samples. The results support investigations of P. vivax infections in archived sera or plasma collections with a high degree of confidence to generate additional data on the neglected P. vivax malaria, and ultimately guide strategies to control the disease.
塞内加尔患者血颗粒和血浆样本中间日疟原虫DNA的分子检测和定量
塞内加尔首次发现间日疟原虫感染病例时,使用的是存档患者的血清,而不是用于疟原虫分子诊断的首选标本血颗粒。目前的研究评估了在血浆中检测间日疟原虫DNA的可靠性,并将其与来自同一患者样本的血液颗粒进行了比较。方法采集塞内加尔Kolda(2015年和2020年)、Tambacounda(2017年和2020年)和Kedougou(2020年)地区发热患者血样616份,采用基于18S ssrrna的巢式PCR方法筛选疟原虫种类组成。从年龄和性别匹配的50例间日疟原虫阳性患者和50例间日疟原虫阴性患者中选择配对的血粒和血浆样本,并进行基于细胞色素b的qPCR,比较两种标本类型间间日疟原虫基因组DNA的检测和定量。该研究报告了研究人群中1.8%和14.77%的单一间日疟原虫感染和混合间日疟原虫感染,配对血粒和血浆样本的间日疟原虫基因组DNA qPCR检测结果高度一致(84%)。重要的是,所有来自血球的间日疟原虫阴性样本也被证实为血浆阴性,血球中的寄生虫血症高于血浆样本。这些结果支持对存档的血清或血浆标本中的间日疟原虫感染进行调查,具有高度的可信度,从而产生关于被忽视的间日疟原虫疟疾的额外数据,并最终指导控制该疾病的战略。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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