Publication: Two-dye based arrayed primer extension for simultaneous multigene detection in lipid metabolism
Issued Date
2015-03-01
Resource Type
ISSN
18733492
00098981
00098981
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2-s2.0-84921532002
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Mahidol University
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SCOPUS
Bibliographic Citation
Clinica Chimica Acta. Vol.442, (2015), 36-43
Suggested Citation
Nutjaree Jeenduang, Sureerut Porntadavity, Markus von Nickisch-Rosenegk, Frank F. Bier, Chamras Promptmas Two-dye based arrayed primer extension for simultaneous multigene detection in lipid metabolism. Clinica Chimica Acta. Vol.442, (2015), 36-43. doi:10.1016/j.cca.2015.01.005 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/35493
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Title
Two-dye based arrayed primer extension for simultaneous multigene detection in lipid metabolism
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Abstract
© 2015 Elsevier B.V. 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 50-150. bp. The total assay including PCR, purification, fragmentation, APEX reaction, and image analysis could be performed in 6. h. 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.
