A multiplex PCR for detection of knockdown resistance mutations, V1016G and F1534C, in pyrethroid-resistant Aedes aegypti

Saingamsook, Jassada; Saeung, Atiporn; Yanola, Jintana; Lumjuan, Nongkran; Walton, Catherine; Somboon, Pradya

doi:10.1186/s13071-017-2416-x

Cited by 39 publications

(51 citation statements)

References 32 publications

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“…To date, more than thirty unique resistance associated mutations or combination of mutations have been detected in pyrethroid and DDT resistant insect populations (Rinkevich et al, 2013). Presence of kdr mutations of mosquito populations have been studied extensively (Singh et al, 2009;Kazanidou et al, 2009;Komagata et al, 2014;Thi et al, 2016;Saingamsook et al, 2017;Nugapola et al, 2017;Hegoda et al, 2017;Saavedra-Rodriguez et al, 2018). Variability in substitution at a single site such as L1014F/H/S/C/W resulting pyrethroids resistance can also be seen.…”

Section: Insensitive Sodium Channel Regulatory Proteinsmentioning

confidence: 99%

Insecticide resistance in mosquitoes: Development, mechanisms and monitoring

et al. 2018

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Mosquito-borne diseases are of major public health concerns, mainly in tropical countries. Control of these diseases is primarily achieved through mosquito control. One of the major challenges encountered by mosquito control programmes is the development of resistance to insecticides by mosquitoes. Increased detoxifying activity through over expression of insect metabolic enzymes and altered target sites with decreased sensitivity to insecticides are the two major mechanisms of resistance. This review article presents our knowledge on development of insecticide resistance and the underlying biochemical and molecular mechanisms of resistance to major classes of insecticides. Techniques available to monitor resistance and to understand the underlying mechanisms are also discussed.

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Section: Insensitive Sodium Channel Regulatory Proteinsmentioning

confidence: 99%

Insecticide resistance in mosquitoes: Development, mechanisms and monitoring

et al. 2018

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“…aegypti mosquitoes from Jakarta (70.70%) [46], Central Java (91.16%) [49], and Denpasar (63.91%) [47]. The 1534C allele frequency in this study (8%) was lower compared to the neighbouring countries of southeast Asia, including Malaysia (40-80%) [45] Vietnam (22%) [43], Myanmar (21%) [43], and Thailand (51%) [64].…”

Section: Discussionmentioning

confidence: 52%

Frequency of kdr mutations in the voltage-sensitive sodium channel (VSSC) gene in Aedes aegypti from Yogyakarta and implications for Wolbachia-infected mosquito trials

Wuliandari

Hoffmann

Tantowijoyo

et al. 2020

Preprint

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Background: In the inner city of Yogyakarta, Indonesia, insecticide resistance is expected in the main dengue vector, Aedes aegypti, because of the intensive local application of pyrethroid insecticides. However, detailed information about the nature of resistance in this species is required to assist the release of Wolbachia mosquitoes in a dengue control program so that we can ensure that insecticide resistance in the strain of Ae. aegypti being released matches that of the background population.Methods: High-resolution melt genotyping was used to screen for kdr mutations associated with pyrethroid resistance in the voltage-sensitive sodium channel (VSSC) gene in Ae. aegypti of some areas in the inner city of Yogyakarta.Results: The results show that the V1016G mutation predominated, with individuals homozygous for the 1016G allele at a frequency of 82.1% and the mutant allele G at a frequency of 92%. Two patterns of co-occurrence of mutations were detected in this study, homozygous individuals V1016G/S989P; and heterozygous individuals V1016G/F1534C/S989P. We found the simultaneous occurrence of kdr mutations V1016G and F1534C at all collection sites, but not within individual mosquitoes. Homozygous mutants at locus 1016 were homozygous wildtype at locus 1534 and vice versa, and heterozygous V1016G were also heterozygous for F1534C. The most common tri-locus genotype co-occurrences were homozygous mutant 1016GG and homozygous wild-type FF1534, combined with homozygous mutant 989PP (GG/FF/PP) at a frequency of 38.28%.Conclusions: Given the relatively small differences in frequency of resistance alleles across the city area, locality variations in resistance should have minor implications for the success of Wolbachia mosquito trials being undertaken in the Yogyakarta area.

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“…[30] for the alleles L/S (TTG/TCG), L/F (TTG/TTT) and L/L (TTG/TTG). Assays have been performed for the detection of V1,016I and F1,534C mutations individually [10, 24], and as multiplex PCR with other mutations such as V1,016G and F1,534C, T1520I and F1,534C [21, 31]. The V410L mutation was first detected in co-occurrence with the V1,016I and F1,534C mutations in Ae.…”

Section: Discussionmentioning

confidence: 99%

“…Recently, the multiplex PCR technique has proven effective in the identification of two kdr mutations (V1,016G and F1,534C) simultaneously in Ae . aegypti [21]. In addition, it is possible to adapt it to different biochemical assays that not only reveal the amplified products optimally, but also contribute to improving the reliability, cost and performance of resistance monitoring [22].…”

Section: Introductionmentioning

confidence: 99%

Multiplex PCR for simultaneous genotyping of kdr mutations V410L, V1,016I and F1,534C inAedes aegypti(L.)

Villanueva-Segura

Ponce‐García

López-Monroy

et al. 2019

Preprint

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BackgroundKnock down resistance (kdr) is the main mechanism that confers resistance to pyrethroids and DDT. This is a product of nonsynonymous mutations in the he voltage-gated sodium channel (VGSC) gene, these mutations produce a change of a single amino acid which reduces the affinity of the target site for the insecticide molecule. In Mexico, V410L, V1,016I and F1,534C mutations are common in pyrethroid-resistant Aedes aegypti (L.) populations.MethodsA multiplex PCR was developed to detect the V410L, V1,016I and F1,534C mutations in Ae. aegypti. The validation of the technique was carried out using wild populations previously characterized for the three mutations through allele-specific PCR (AS-PCR) and with different levels of genotypic frequencies.ResultsThe standardized protocol for multiplex endpoint PCR was highly effective in detecting 12 genotypes in five wild Ae. aegypti populations from Mexico. A complete concordance with AS-PCR was found for the simultaneous detection of the three kdr mutations.ConclusionsOur diagnostic method is highly effective for the simultaneous detection of V410L, V1,016I and F1,534C, when they co-occur. This technique represents a viable alternative to complement and strengthen current monitoring and resistance management strategies against Ae. aegypti.

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A multiplex PCR for detection of knockdown resistance mutations, V1016G and F1534C, in pyrethroid-resistant Aedes aegypti

Cited by 39 publications

References 32 publications

Insecticide resistance in mosquitoes: Development, mechanisms and monitoring

Insecticide resistance in mosquitoes: Development, mechanisms and monitoring

Frequency of kdr mutations in the voltage-sensitive sodium channel (VSSC) gene in Aedes aegypti from Yogyakarta and implications for Wolbachia-infected mosquito trials

Multiplex PCR for simultaneous genotyping of kdr mutations V410L, V1,016I and F1,534C inAedes aegypti(L.)

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