Application of loop-mediated isothermal amplification combined with lateral flow assay visualization of Plasmodium falciparum kelch 13 C580Y mutation for artemisinin resistance detection in clinical samples

Sanmoung, Wannida; Sawangjaroen, Nongyao; Jitueakul, Suwannee; Buncherd, Hansuk; Tun, Aung Win; Thanapongpichat, Supinya; Imwong, Mallika

doi:10.1016/j.actatropica.2023.106998

Cited by 3 publications

(3 citation statements)

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“…falciparum drug resistance to artemisinin and its derivatives (e.g., artesunate). These include the use of loop-mediated isothermal amplification (LAMP), ,, a method based on amplification of the readout signal with DNA primers. In the current study, we show, for the first time, a simple sensing system for detecting this SNP without the need of amplifying the signal.…”

Section: Resultsmentioning

confidence: 99%

“…In recent years, several diagnostic approaches have been devised to allow a relatively simple approach to identify the C580Y SNP in the K13 gene that is associated with P. falciparum drug resistance to artemisinin and its derivatives (e.g., artesunate). These include the use of loop-mediated isothermal amplification (LAMP), 16 , 17 , 19 a method based on amplification of the readout signal with DNA primers. In the current study, we show, for the first time, a simple sensing system for detecting this SNP without the need of amplifying the signal.…”

Section: Resultsmentioning

confidence: 99%

“…While there have been attempts to develop SNP detection methods for point-of-care treatment, the current detection methods mainly rely on PCR-based technologies − and isothermal amplification techniques. These methods, such as loop-mediated isothermal amplification (LAMP), − drop-digital PCR and molecular beacon PCR, single-nucleotide primer extension (SNPE), and fluorescence resonance energy transfer-melting curve analysis (FRET-MCA), are highly sensitive but are often time-consuming and may require expensive equipment, sensitive materials (polymerases), and skilled personnel. These requirements pose challenges in resource-limited malaria-endemic countries, where such facilities and expertise may not be readily available.…”

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confidence: 99%

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A Biotinylated cpFIT-PNA Platform for the Facile Detection of Drug Resistance to Artemisinin in Plasmodium falciparum

Tepper,

Appella,

Zheng

et al. 2024

ACS Sens.

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The evolution of drug resistance to many antimalarial drugs in the lethal strain of malaria (Plasmodium falciparum) has been a great concern over the past 50 years. Among these drugs, artemisinin has become less effective for treating malaria. Indeed, several P. falciparum variants have become resistant to this drug, as elucidated by specific mutations in the pfK13 gene. This study presents the development of a diagnostic kit for the detection of a common point mutation in the pfK13 gene of P. falciparum, namely, the C580Y point mutation. FIT-PNAs (forced-intercalation peptide nucleic acid) are DNA mimics that serve as RNA sensors that fluoresce upon hybridization to their complementary RNA. Herein, FIT-PNAs were designed to sense the C580Y single nucleotide polymorphism (SNP) and were conjugated to biotin in order to bind these molecules to streptavidin-coated plates. Initial studies with synthetic RNA were conducted to optimize the sensing system. In addition, cyclopentane-modified PNA monomers (cpPNAs) were introduced to improve FIT-PNA sensing. Lastly, total RNA was isolated from red blood cells infected with P. falciparum (WT strain – NF54-WT or mutant strain – NF54-C580Y). Streptavidin plates loaded with either FIT-PNA or cpFIT-PNA were incubated with the total RNA. A significant difference in fluorescence for mutant vs WT total RNA was found only for the cpFIT-PNA probe. In summary, this study paves the way for a simple diagnostic kit for monitoring artemisinin drug resistance that may be easily adapted to malaria endemic regions.

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Section: Resultsmentioning

confidence: 99%