Detection of the East and West African kdr mutation in Anopheles gambiae and Anopheles arabiensis from Uganda using a new assay based on FRET/Melt Curve analysis

Malaria Journal
Katrijn VerhaeghenMarc Coosemans

Abstract

Appropriate monitoring of vector resistance to insecticides is an integral component of planning and evaluation of insecticide use in malaria control programmes. The malaria vectors Anopheles gambiae s.s. and Anopheles arabiensis have developed resistance to pyrethroid insecticides as a result of a mechanism conferring reduced nervous system sensitivity, better known as knockdown resistance (kdr). In An. gambiae s.s. and An. arabiensis, two different substitutions in the para-type sodium channel, a L1014F substitution common in West Africa and a L1014S replacement found in Kenya, are linked with kdr. Two different allele-specific polymerase chain reactions (AS-PCR) are needed to detect these known kdr mutations. However, these AS-PCR assays rely on a single nucleotide polymorphism mismatch, which can result in unreliable results. Here, a new assay for the detection of knockdown resistance in An. gambiae s.s. and An. arabiensis based on Fluorescence Resonance Energy Transfer/Melt Curve analysis (FRET/MCA) is presented and compared with the existing assays. The new FRET/MCA method has the important advantage of detecting both kdr alleles in one assay. Moreover, results show that the FRET/MCA is more reliable and more sensitive th...Continue Reading

Citations

Sep 18, 2008·Malaria Journal·Frederic PagèsCheikh Sokhna
Nov 1, 2011·BMC Research Notes·Philippe NwaneFrédéric Simard
Mar 30, 2010·The American Journal of Tropical Medicine and Hygiene·Katrijn VerhaeghenMarc Coosemans
Jul 3, 2010·The American Journal of Tropical Medicine and Hygiene·Delenasaw YewhalawNiko Speybroeck
Jan 11, 2012·The American Journal of Tropical Medicine and Hygiene·Cara N Henry-HalldinPeter A Zimmerman
Dec 12, 2012·Transactions of the Royal Society of Tropical Medicine and Hygiene·Jean Pierre Basilua KanzaFrancis Watsenga Tezzo
Feb 27, 2013·The American Journal of Tropical Medicine and Hygiene·Michael R ReddyMichel A Slotman
May 10, 2007·Insect Molecular Biology·W C Black, J G Vontas
Jul 8, 2016·Ciência & saúde coletiva·Guilherme Liberato da SilvaOnilda Santos da Silva
Sep 25, 2017·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Olivier Gnankiné, Imaël Henri Nestor Bassolé
Nov 6, 2008·Tropical Medicine & International Health : TM & IH·Natacha ProtopopoffMarc Coosemans

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Datasets Mentioned

BETA
DQ263749
DQ263748

Methods Mentioned

BETA
Assay
Fluorescence
enzyme linked immunosorbent assay
PCR
FRET
ELISA
genotyping

Software Mentioned

Meltcalc
[UNK] iQ Optical system

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