title: Two-dye based arrayed primer extension for simultaneous multigene detection in lipid metabolism.
abstract: BACKGROUND
  Cardiovascular disease (CVD) is one of the major causes of death worldwide. Numerous genetic risk factors in lipid metabolism, including mutations of LDLR, APOB, and PCSK9, as well as polymorphisms of CETP and APOE, have been found to associate with CVD.
  
  
  METHODS
  In this study, a two-dye based arrayed primer extension (APEX) microarray assay for simultaneous multigene (LDLR, APOB, PCSK9, CETP, and APOE) detection was developed. The DNA templates, originating from 1 DNA sample of known genotype and 7 blind DNA samples, were amplified by uniplex PCR.
  
  
  RESULTS
  Optimized conditions for the APEX reaction were determined to include a hybridization temperature of 55°C and a DNA template size of 50-150bp. The total assay including PCR, purification, fragmentation, APEX reaction, and image analysis could be performed in 6h. In total, 48 genotypes were identified among 8 individual DNA samples by APEX analysis.
  
  
  CONCLUSIONS
  The data suggest that this APEX microarray offers a robust, fast, and versatile option for screening these genotypes in hypercholesterolemia patients.
authors.affiliation: School of Allied Health Science and Public Health, Walailak University, Nakhon Si Thammarat 80161, Thailand; Department of Nanobiotechnology and Nanomedicine, Fraunhofer Institute for Biomedical Engineering (IBMT), Potsdam-Golm 14476, Germany.
authors.forename: Nutjaree
authors.initials: N
authors.lastname: Jeenduang
authors.affiliation: Department of Clinical Chemistry, Faculty of Medical Technology, Mahidol University, Nakhon Pathom 73170, Thailand.
authors.forename: Sureerut
authors.initials: S
authors.lastname: Porntadavity
authors.affiliation: Department of Nanobiotechnology and Nanomedicine, Fraunhofer Institute for Biomedical Engineering (IBMT), Potsdam-Golm 14476, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: von Nickisch-Rosenegk
authors.affiliation: Department of Nanobiotechnology and Nanomedicine, Fraunhofer Institute for Biomedical Engineering (IBMT), Potsdam-Golm 14476, Germany.
authors.forename: Frank F
authors.initials: FF
authors.lastname: Bier
authors.affiliation: Department of Biomedical Engineering, Faculty of Engineering, Mahidol University, Nakhon Pathom 73170, Thailand. Electronic address: chamras.pro@mahidol.ac.th.
authors.forename: Chamras
authors.initials: C
authors.lastname: Promptmas
mesh_terms: D004396:Coloring Agents / Q000378:metabolism*; D017931:DNA Primers / Q000235:genetics*; D005838:Genotype; D006801:Humans; D050356:Lipid Metabolism / Q000235:genetics*; D005810:Multigene Family / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction; D013696:Temperature; D013997:Time Factors
chemical_list: D004396:Coloring Agents; D017931:DNA Primers
keywords: Arrayed primer extension (APEX); Genotyping; Lipid metabolism; Microarray