Development a point-of-care lateral flow test for artemisinin-resistant malaria

Artemisinin-based combination therapy is currently recommended by WHO for treatment of
uncomplicated falciparum malaria. However, artemisinin resistance is threatening the effectiveness
of these antimalarials and has spread rapidly in South-East Asia [1]. Should artemisinin-resistant
malaria (ARM) spread from this region to Africa, as did resistance to older antimalarials, much of the
progress made in reducing the prevalence of malaria could be reversed.
While the mechanism of ARM is still only partially understood, multiple single-nucleotide
polymorphisms in the propeller domain of a kelch gene, located on chromosome 13, have been
shown to be associated with artemisinin resistance [2]. Two single nucleotide polymorphisms,
C580Y and R539T, are the most prevalent SNPs associated with ARM and account for 85% in SouthEast Asia, with some areas having a prevalence as high as 90% [3].
Since the discovery of the molecular markers for ARM, nucleic-acid based tests such as polymerasechain reaction (PCR) have been used to test for ARM. However PCR-based techniques require
complex apparatus, trained personnel and external power, and therefore are not readily available in
resource-poor settings. This method is useful for epidemiological studies but does not allow for
results to be used in case-management as the molecular testing is conducted long after the
collection of blood from the patient.
To overcome these challenges, other techniques such as loop-mediated isothermal amplification
(LAMP)-based methods have been developed. LAMP has many advantages due to its low cost and
simplicity, and is increasingly used as point-of-care tests by clinicians for other infectious diseases. It
has been shown to be user-friendly and cost-effective [4]. Microfluidics
To date there is no point-of-care (POC) test available to detect ARM. A POC test for AMR in malaria
endemic areas could help to inform clinicians about appropriate case management for patients.
Currently alternative drug regimens such as the use of partner drugs and longer treatments are
being evaluated for ARM, however this can lead to drug toxicity and multidrug-resistant malaria [5].
POC tests for ARM would therefore enable clinicians to provide appropriate treatment and limit the
overuse of existing antimalarials. POC tests would also provide real time data for epidemiological
studies.
Objectives
The main objective of this project is to develop a new low cost test that will combine microfluidics
and finger prick sampling to enable for the first time the testing of ARM at the POC. The test would
use LAMP to test for the most common SNPs conferring ARM and would use lateral flow and
microfluidic technology. The test should have sensitivity and specificity as close as possible to PCR,
while being suitable for use in low-resource environments, with the minimal requirements for
additional equipment.