Rapid low-resource detection of Plasmodium falciparum in infected Anopheles mosquitoes

Leon E. Hugo; Karla van Huyssteen; Olamide Oloniniyi; Laura Donnelly; Anna Conn; Katharine A. Collins; Hayley Mitchell; James S. McCarthy; Joanne Macdonald

doi:10.3389/fitd.2024.1287025

Back

Rapid low-resource detection of Plasmodium falciparum in infected Anopheles mosquitoes

Journal article

Open access

Peer reviewed

Rapid low-resource detection of Plasmodium falciparum in infected Anopheles mosquitoes

Leon E. Hugo, Karla van Huyssteen, Olamide Oloniniyi, Laura Donnelly, Anna Conn, Katharine A. Collins, Hayley Mitchell, James S. McCarthy and Joanne Macdonald

Frontiers in Tropical Diseases, Vol.5, pp.1-10

2024

DOI: https://doi.org/10.3389/fitd.2024.1287025

Files and links (1)

pdf

fitd-05-12870252.27 MBDownload View

Published VersionCC BY V4.0, Open Access

Abstract

isothermal (DNA) amplification

rapid test

malaria - mosquito vectors

rapid sample preparation

nucleic acid lateral flow

Plasmodium falciparum

Vector surveillance of Plasmodium falciparum is critical for monitoring and reducing one of the most severe forms of malaria, which causes high morbidity and mortality in children under five and pregnant women. Here we developed a rapid and highly sensitive test for the detection of P. falciparum (Pf) -infected mosquitoes (Rapid Pf test), with high suitability for low-resource vector surveillance implementation. The Rapid Pf test had similar analytical sensitivity to laboratory-based tests, detecting down to 4 copies/μL of a 18S rRNA DNA standard. In addition, the Rapid Pf test could be completed in less than 30 minutes, and only required a liquid sample preparation reagent, pestle, tube, and 39°C heating block for operation, indicating amenability for low-resource implementation. Diagnostic testing was performed using Anopheles stephensi mosquitoes, either uninfected, or fed with P. falciparum gametocyte cultures. These P. falciparum fed mosquitoes were determined to have 79% infection prevalence based on parallel microscopy and qPCR testing on a subset of 19 mosquitoes. However, our Rapid Pf test determined a 90% positive test rate when testing individual infected mosquitoes (n=30), and did not detect 40 uninfected mosquitoes regardless of blood-fed status (n=40), suggesting the true prevalence of infection in the mosquitoes may have been higher than calculated by qPCR and microscopy. The Rapid Pf test was demonstrated to detect infection in individual mosquitoes (both fresh and frozen/thawed), as well as pools of 1 infected mosquito mixed with 19 known uninfected mosquitoes, and individual mosquitoes left in traps for up to 8 days. After testing on infected and uninfected mosquitoes (n=148) the Rapid Pf test was conservatively estimated to achieve 100% diagnostic sensitivity (95% confidence interval, CI: 91%-100%) and 97% diagnostic specificity (CI: 92%-99%) compared to the estimated prevalence from combined microscopy and qPCR results. These results indicate the Rapid Pf test could provide a highly effective tool for weekly surveillance of infected mosquitoes, to assist with P. falciparum monitoring and intervention studies.

Details

Title: Rapid low-resource detection of Plasmodium falciparum in infected Anopheles mosquitoes
Authors: Leon E. Hugo (Author) - QIMR Berghofer Medical Research Institute
Karla van Huyssteen (Author) - University of the Sunshine Coast, Queensland, Centre for Bioinnovation
Olamide Oloniniyi (Author) - University of the Sunshine Coast, Queensland, Centre for Bioinnovation
Laura Donnelly (Author) - University of the Sunshine Coast, Queensland, Centre for Bioinnovation
Anna Conn (Author) - University of the Sunshine Coast, Queensland, Centre for Bioinnovation
Katharine A. Collins (Author) - QIMR Berghofer Medical Research Institute
Hayley Mitchell (Author) - QIMR Berghofer Medical Research Institute
James S. McCarthy (Author) - QIMR Berghofer Medical Research Institute
Joanne Macdonald (Corresponding Author) - University of the Sunshine Coast, Queensland, Centre for Bioinnovation
Publication details: Frontiers in Tropical Diseases, Vol.5, pp.1-10
Publisher: Frontiers Research Foundation
Date published: 2024
DOI: 10.3389/fitd.2024.1287025
ISSN: 2673-7515
Copyright note: © 2024 Hugo, van Huyssteen, Oloniniyi, Donnelly, Conn, Collins, Mitchell, McCarthy and Macdonald. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
Grant note: Bill and Melinda Gates Foundation [OPP1140133]
Organisation Unit: UniSC Clinical Trials Centre; School of Science, Technology and Engineering; Centre for Bioinnovation
Language: English
Record Identifier: 99996896102621
Output Type: Journal article

Metrics

4 File views/ downloads

155 Record Views