Evidence map›Paper›PMID 25245258›Full record

ArticleMalaria journal2014

Development and application of an AllGlo probe-based qPCR assay for detecting knockdown resistance (kdr) mutations in Anopheles sinensis.

Liang Bai, Guo-ding Zhu, Hua-yun Zhou, Jian-xia Tang, Ju-lin Li, Sui Xu, Mei-hua Zhang, Li-nong Yao, Guang-quan Huang, Yong-bin Wang and 4 more

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Article in Malaria journal, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.

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5citing papers in PubMed
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1 · What the graph read from it

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3 · Its place in the literature

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5 citing papers in PubMed.

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4 · The record

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5 · Who and what money

Authors and funding

14 authors.

Liang Bai
Guo-ding Zhu
Hua-yun Zhou
Jian-xia Tang
Ju-lin Li
Sui Xu
Mei-hua Zhang
Li-nong Yao
Guang-quan Huang
Yong-bin Wang
Hong-wei Zhang
Si-bao Wang
Jun CaoKey Laboratory of Parasitic Disease Control and Prevention (Ministry of Health), and Jiangsu Provincial Key Laboratory of Parasite Molecular Biology, Jiangsu Institute of Parasitic Diseases, Wuxi, Jiangsu Province, People's Republic of China. caojuncn@hotmail.com.
Qi Gao

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

backgroundAnopheles sinensis is one of the most important malaria vectors in China and other Southeast Asian countries. High levels of resistance have been reported in this species due to the long-term use of insecticides, especially pyrethroids, for public health and agricultural purposes. Knockdown resistance (kdr) caused by a single base pair mutation in the gene encoding the sodium channel is strongly associated with pyrethroid insecticide resistance in many Anopheles mosquitoes. There are few methods currently available for detecting kdr mutations in An. sinensis.

methodsA novel AllGlo probe-based qPCR (AllGlo-qPCR) method was developed to screen for the predominant kdr mutations in An. sinensis mosquitoes from the Jiangsu Province. The results from AllGlo-qPCR, allele-specific PCR (AS-PCR), and TaqMan-MGB probe-based qPCR (TaqMan-qPCR) were compared. A comparative analysis of the equipment required, ease of use and cost of the available methods was also performed. Finally, the AllGlo-qPCR method was used to detect the frequencies of kdr mutations from the other four provinces in central China.

resultsSix kdr genotypes were detected in An. sinensis from the Jiangsu Province by DNA sequencing. The AllGlo-qPCR method detected all of the kdr genotypes with a high level of accuracy (97% sensitivity and 98% specificity). AllGlo-qPCR correctly determined the kdr genotypes of 98.73% of 158 An. sinensis samples, whereas TaqMan-qPCR and AS-PCR correctly identified 96.84% and 88.61% of mutations, respectively. Furthermore, the AllGlo-qPCR method is simpler to perform, requires less equipment, and exhibits a moderate expense cost comparing with the other tested methods of kdr mutation detection. Samples collected from four of the other provinces in central China showed a high frequency of kdr mutation in An. sinensis, as detected by the established AllGlo-qPCR method.

conclusionThe novel AllGlo-qPCR method developed for kdr mutation detection in An. sinensis exhibits greater specificity and sensitivity than currently available methods and is more cost-effective; therefore, it represents a useful tool for entomological surveillance.

Indexed as

Molecular Probe TechniquesAnimalsAnophelesBase SequenceChinaGenotypeInsecticide ResistanceMolecular Sequence DataMutationPolymerase Chain ReactionSensitivity and Specificity

Identifiers

PMID25245258
PMCPMC4182860

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