Targeted deep amplicon sequencing of kelch 13 and cytochrome b in Plasmodium falciparum isolates from an endemic African country using the Malaria Resistance Surveillance (MaRS) protocol

利用疟疾耐药性监测(MaRS)方案,对来自非洲疟疾流行国家的恶性疟原虫分离株中的Kelch 13和细胞色素b进行靶向深度扩增子测序。

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作者:Mariangela L'Episcopia ,Julia Kelley ,Dhruviben Patel ,Sarah Schmedes ,Shashidahar Ravishankar ,Michela Menegon ,Edvige Perrotti ,Abduselam M Nurahmed ,Albadawi A Talha ,Bakri Y Nour ,Naomi Lucchi ,Carlo Severini ,Eldin Talundzic

Abstract

Background: Routine molecular surveillance for imported drug-resistant malaria parasites to the USA and European Union is an important public health activity. The obtained molecular data are used to help keep chemoprophylaxis and treatment guidelines up to date for persons traveling to malaria endemic countries. Recent advances in next-generation sequencing (NGS) technologies provide a new and effective way of tracking malaria drug-resistant parasites. Methods: As part of a technology transfer arrangement between the CDC Malaria Branch and the Istituto Superiore di Sanità (ISS), Rome, Italy, the recently described Malaria Resistance Surveillance (MaRS) protocol was used to genotype 148 Plasmodium falciparum isolates from Eritrea for kelch 13 (k13) and cytochrome b (cytb) genes, molecular markers associated with resistance to artemisinin (ART) and atovaquone/proguanil (AP), respectively. Results: Spanning the full-length k13 gene, seven non-synonymous single nucleotide polymorphisms (SNPs) were found (K189N, K189T, E208K, D281V, E401Q, R622I and T535M), of which none have been associated with artemisinin resistance. No mutations were found in cytochrome b. Conclusion: All patients successfully genotyped carried parasites susceptible to ART and AP treatment. Future studies between CDC Malaria Branch and ISS are planned to expand the MaRS system, including data sharing, in an effort to maintain up to date treatment guidelines for travelers to malaria endemic countries. Keywords: Drug resistance; Molecular surveillance; Next-generation sequencing; Plasmodium falciparum.

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