University of Basel, Petersplatz 1, 4001 Basel, Switzerland.
Department of Medical Parasitology and Infection Biology, Swiss Tropical and Public Health Institute, Kreuzstrasse 2, 4123 Allschwil, Switzerland.
J Antimicrob Chemother. 2024 May 2;79(5):987-996. doi: 10.1093/jac/dkae060.
BACKGROUND: The emergence of drug-resistant clones of Plasmodium falciparum is a major public health concern, and the ability to detect and track the spread of these clones is crucial for effective malaria control and treatment. However, in endemic settings, malaria infected people often carry multiple P. falciparum clones simultaneously making it likely to miss drug-resistant clones using traditional molecular typing methods. OBJECTIVES: Our goal was to develop a bioinformatics pipeline for compositional profiling in multiclonal P. falciparum samples, sequenced using the Oxford Nanopore Technologies MinION platform. METHODS: We developed the 'Finding P. falciparum haplotypes with resistance mutations in polyclonal infections' (PHARE) pipeline using existing bioinformatics tools and custom scripts written in python. PHARE was validated on three control datasets containing P. falciparum DNA of four laboratory strains at varying mixing ratios. Additionally, the pipeline was tested on clinical samples from children admitted to a paediatric hospital in the Central African Republic. RESULTS: The PHARE pipeline achieved high recall and accuracy rates in all control datasets. The pipeline can be used on any gene and was tested with amplicons of the P. falciparum drug resistance marker genes pfdhps, pfdhfr and pfK13. CONCLUSIONS: The PHARE pipeline helps to provide a more complete picture of drug resistance in the circulating P. falciparum population and can help to guide treatment recommendations. PHARE is freely available under the GNU Lesser General Public License v.3.0 on GitHub: https://github.com/Fippu/PHARE.
背景:疟原虫耐药克隆的出现是一个主要的公共卫生关注点,能够检测和跟踪这些克隆的传播对于有效控制和治疗疟疾至关重要。然而,在流行地区,感染疟疾的人通常同时携带多种疟原虫克隆,因此使用传统的分子分型方法可能会错过耐药克隆。
目的:我们的目标是开发一种针对多克隆疟原虫样本的组成分析生物信息学管道,这些样本是使用牛津纳米孔技术 MinION 平台进行测序的。
方法:我们使用现有的生物信息学工具和用 python 编写的自定义脚本开发了“在多克隆感染中寻找具有耐药突变的疟原虫单倍型”(PHARE)管道。PHARE 在三个包含四种实验室菌株在不同混合比例的疟原虫 DNA 的对照数据集上进行了验证。此外,该管道还在中非共和国一家儿科医院收治的儿童的临床样本上进行了测试。
结果:PHARE 管道在所有对照数据集上都实现了高召回率和准确率。该管道可用于任何基因,并已在疟原虫耐药标记基因 pfdhps、pfdhfr 和 pfK13 的扩增子上进行了测试。
结论:PHARE 管道有助于更全面地了解循环疟原虫群体中的耐药情况,并有助于指导治疗建议。PHARE 可在 GitHub 上根据 GNU Lesser General Public License v.3.0 免费获得:https://github.com/Fippu/PHARE。
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