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Development of a low‐density DNA microarray for diagnosis of target‐site mutations of pyrethroid and organophosphate resistance mutations in the whitefly Bemisia tabaci
Author(s) -
Chung InHyuk,
Kang Soyoung,
Kim Ye Rim,
Kim JiHoon,
Jung Jin Wook,
Lee Seunghwan,
Lee Si Hyeock,
Hwang Seung Yong
Publication year - 2011
Publication title -
pest management science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.296
H-Index - 125
eISSN - 1526-4998
pISSN - 1526-498X
DOI - 10.1002/ps.2209
Subject(s) - biology , multiplex , genetics , microarray , dna microarray , gene , mutation , genotype , microbiology and biotechnology , gene expression
BACKGROUND: Rapid and accurate detection of mutations related to insecticide resistance is essential for development of resistance management strategies to support sustainable agriculture. The M918V, L925I and T929V mutations of the voltage‐gated sodium channel gene ( vgsc ) and the F392W mutation of the acetylcholinesterase I gene ( ace1 ) are reportedly associated with resistance to pyrethroids and organophosphates, respectively, in Bemisia tabaci . In order to detect known base substitutions in the ace1 and vgsc genes, a low‐density microarray with an allele‐specific probe was developed. RESULTS: Specific regions of the ace1 and vgsc gene mutations were amplified by multiplex asymmetrical PCR using Cy3‐labelled primers, and then the PCR products were hybridised on the microarray. After analysing the probe signal data, the microarray containing 12 allele‐specific probes produced a unique pattern of probe signals for field DNA samples of B. tabaci. To determine the optimal cut‐off value of each probe, receiver operating characteristic (ROC) curve analysis was conducted using SPSS. Among 60 individual samples, microarray data for 57 samples were consistent with direct sequencing data. CONCLUSION: Although many molecular detection methods have been employed to monitor insecticide resistance, the present microarray provides rapid and accurate identification of target mutations in B. tabaci for resistance management. Copyright © 2011 Society of Chemical Industry