Molecular Surveillance of Malaria Using the PF AmpliSeq Custom Assay for Plasmodium falciparum Parasites from Dried Blood Spot DNA Isolates from Peru

Autor: Johanna Kattenberg, Norbert van Dijk, Carlos Fernández-Miñope, Pieter Guetens, Mathijs Mutsaers, Dionicia Gamboa, Anna Rosanas-Urgell
Jazyk: angličtina
Rok vydání: 2023
Předmět:
Zdroj: Bio-Protocol, Vol 13, Iss 5 (2023)
Druh dokumentu: article
ISSN: 2331-8325
DOI: 10.21769/BioProtoc.4621
Popis: Malaria molecular surveillance has great potential to support national malaria control programs (NMCPs), informing policy for its control and elimination. Here, we present a new three-day workflow for targeted resequencing of markers in 13 resistance-associated genes, histidine rich protein 2 and 3 (hrp2&3), a country (Peru)-specific 28 SNP-barcode for population genetic analysis, and apical membrane antigen 1 (ama1), using Illumina short-read sequencing technology. The assay applies a multiplex PCR approach to amplify all genomic regions of interest in a rapid and easily standardizable procedure and allows simultaneous amplification of a high number of targets at once, therefore having great potential for implementation into routine surveillance practice by NMCPs. The assay can be performed on routinely collected filter paper blood spots and can be easily adapted to different regions to investigate either regional trends or in-country epidemiological changes.
Databáze: Directory of Open Access Journals