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Molecular detection and quantification of Plasmodium falciparum gametocytes carriage in used RDTs in malaria elimination settings in northern Senegal
Malaria Journal ( IF 3 ) Pub Date : 2020-03-30 , DOI: 10.1186/s12936-020-03204-w
Kiswendsida Thierry Guiguemde 1 , Yakou Dieye 2 , Aminata Collé Lô 1 , Magatte Ndiaye 1 , Aminata Lam 1 , Isaac Akhénaton Manga 1 , Gnagna Dieng Sow 2 , Moussa Diop 2 , Tamba Souané 2 , Marie Pièrre Diouf 1 , Roger Clément Kouly Tine 1 , Babacar Faye 1
Affiliation  

Malaria surveillance requires powerful tools and strategies to achieve malaria elimination. Rapid diagnostic tests for malaria (RDTs) are easily deployed on a large scale and are helpful sources of parasite DNA. The application of sensitive molecular techniques to these RDTs is a modern tool for improving malaria case detection and drug resistance surveillance. Several studies have made it possible to extract the DNA of Plasmodium falciparum from RDTs. The knowledge of gametocyte carriage in the population is important to better assess the level of parasite transmission in elimination settings. The aim of this study was to detect P. falciparum gametocytes from used RDTs by quantitative PCR for molecular monitoring of malaria transmission. DNA was extracted from 303 RDT devices (SD Bioline Malaria Pf) using the Chelex-100 protocol. qPCR was performed in a 20 μL reaction to detect and quantify transcripts of the pfs25 gene. The cycle threshold (Ct) was determined by the emission fluorescence corresponding to the initial amount of amplified DNA. The study found an overall prevalence of 53.47% with an average Ct of 32.12 ± 4.28 cycles. In 2018, the prevalence of gametocytes was higher in the Ranérou district (76.24%) than in the Saint-Louis district (67.33%) where an increase in the number of gametocyte carriers in 2018 was noted, in comparison with 2017. RDTs are a good source of DNA for molecular monitoring of gametocyte carriage. This method is a simple and effective tool to better understand the level of malaria transmission with a view to elimination.

中文翻译:

塞内加尔北部消除疟疾环境中使用的 RDT 中恶性疟原虫配子体携带的分子检测和定量

疟疾监测需要强大的工具和战略来实现消除疟疾。疟疾快速诊断测试 (RDT) 很容易大规模部署,并且是寄生虫 DNA 的有用来源。将敏感分子技术应用于这些 RDT 是改善疟疾病例检测和耐药性监测的现代工具。多项研究使得从 RDT 中提取恶性疟原虫 DNA 成为可能。了解人群中配子体携带情况对于更好地评估消除环境中寄生虫传播的水平非常重要。本研究的目的是通过定量 PCR 检测使用过的 RDT 中的恶性疟原虫配子细胞,用于疟疾传播的分子监测。使用 Chelex-100 方案从 303 RDT 设备 (SD Bioline Malaria Pf) 中提取 DNA。在 20 μL 反应中进行 qPCR,以检测和定量 pfs25 基因的转录本。循环阈值 (Ct) 由对应于扩增 DNA 初始量的发射荧光确定。研究发现总体患病率为 53.47%,平均 Ct 为 32.12 ± 4.28 个周期。2018 年,Ranérou 区的配子体患病率 (76.24%) 高于圣路易斯区 (67.33%),与 2017 年相比,2018 年配子体携带者数量有所增加。用于分子监测配子体携带的 DNA 的良好来源。该方法是一种简单有效的工具,可以更好地了解疟疾传播程度,以期消除疟疾。循环阈值 (Ct) 由对应于扩增 DNA 初始量的发射荧光确定。研究发现总体患病率为 53.47%,平均 Ct 为 32.12 ± 4.28 个周期。2018 年,Ranérou 区的配子体患病率 (76.24%) 高于圣路易斯区 (67.33%),与 2017 年相比,2018 年配子体携带者数量有所增加。用于分子监测配子体携带的 DNA 的良好来源。该方法是一种简单有效的工具,可以更好地了解疟疾传播程度,以期消除疟疾。循环阈值 (Ct) 由对应于扩增 DNA 初始量的发射荧光确定。研究发现总体患病率为 53.47%,平均 Ct 为 32.12 ± 4.28 个周期。2018 年,Ranérou 区的配子体患病率 (76.24%) 高于圣路易斯区 (67.33%),与 2017 年相比,2018 年配子体携带者数量有所增加。用于分子监测配子体携带的 DNA 的良好来源。该方法是一种简单有效的工具,可以更好地了解疟疾传播程度,以期消除疟疾。Ranérou 区的配子体患病率 (76.24%) 高于圣路易斯区 (67.33%),与 2017 年相比,2018 年配子体携带者数量有所增加。RDT 是配子体携带者的良好来源。用于配子体运输分子监测的 DNA。该方法是一种简单有效的工具,可以更好地了解疟疾传播程度,以期消除疟疾。Ranérou 区的配子体患病率 (76.24%) 高于圣路易斯区 (67.33%),与 2017 年相比,2018 年配子体携带者数量有所增加。RDT 是配子体携带者的良好来源。用于配子体运输分子监测的 DNA。该方法是一种简单有效的工具,可以更好地了解疟疾传播程度,以期消除疟疾。
更新日期:2020-04-22
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