title: Development of a low-density DNA microarray for diagnosis of target-site mutations of pyrethroid and organophosphate resistance mutations in the whitefly Bemisia tabaci.
abstract: 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Biochemistry, Hanyang University and GenoCheck Co., Ltd, Sa-Dong, Sangrok-Gu, Ansan, Gyeonggi-Do, Korea.
authors.forename: In-Hyuk
authors.initials: IH
authors.lastname: Chung
authors.forename: Soyoung
authors.initials: S
authors.lastname: Kang
authors.forename: Ye Rim
authors.initials: YR
authors.lastname: Kim
authors.forename: Ji-Hoon
authors.initials: JH
authors.lastname: Kim
authors.forename: Jin Wook
authors.initials: JW
authors.lastname: Jung
authors.forename: Seunghwan
authors.initials: S
authors.lastname: Lee
authors.forename: Si Hyeock
authors.initials: SH
authors.lastname: Lee
authors.forename: Seung Yong
authors.initials: SY
authors.lastname: Hwang
mesh_terms: D000110:Acetylcholinesterase / Q000235:genetics; D000483:Alleles; D000818:Animals; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D015342:DNA Probes / Q000235:genetics; D017344:Genes, Insect; D006430:Hemiptera / Q000187:drug effects / Q000235:genetics*; D007305:Insecticide Resistance; D007306:Insecticides / Q000494:pharmacology*; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D010755:Organophosphates / Q000494:pharmacology*; D011722:Pyrethrins / Q000494:pharmacology*; D012372:ROC Curve
chemical_list: D017931:DNA Primers; D015342:DNA Probes; D007306:Insecticides; D010755:Organophosphates; D011722:Pyrethrins; D000110:Acetylcholinesterase