TY - JOUR
T1 - Time-saving multiplex detection of single nucleotide polymorphisms by ultrasensitive DNA microarray
AU - Ichikawa, Makiko
AU - Miwa, Keishi
AU - Yamasaki, Tomo
AU - Nakagawa, Izumi
AU - Takizawa, Satoko
AU - Masuda, Satohiro
AU - Inui, Ken Ichi
N1 - Funding Information:
This work was supported in part by New Energy and Industrial Technology Development Organization (NEDO) through ‘Development of Practical Biological Tools’.
Copyright:
Copyright 2011 Elsevier B.V., All rights reserved.
PY - 2010/11
Y1 - 2010/11
N2 - Rapid and multiplex detection system using an ultrasensitive DNA microarray was developed and utilized for the analysis of six pharmacokinetically relevant single nucleotide polymorphisms (SNPs) (MDR1-C1236T, MDR1-G2677TA, MDR1-C3435T, CYP3A5-A6986G, CYP2C19-G681A, CYP2C19-G636A) from blood samples derived from liver transplant patients. The SNP detection system is comprised of three processes: multiplex PCR, single base extension with fluorescently labelled di-deoxy-nucleotides and detection by DNA microarray. The entire workflow of this system completes within 5 h. The final genotype call was obtained statistically by Mahalanobis distance which was calculated from the bi-coloured fluorescent signals detected by the microarray. In order to detect the six SNPs, this system required only 50 copies of genomic DNA, and the obtained detection calls completely matched with the results by the sequencing-based genotyping method. With the high sensitivity and rapid processing, our SNP detection system utilizing ultrasensitive microarray is a promising device applicable for diagnostic utility.
AB - Rapid and multiplex detection system using an ultrasensitive DNA microarray was developed and utilized for the analysis of six pharmacokinetically relevant single nucleotide polymorphisms (SNPs) (MDR1-C1236T, MDR1-G2677TA, MDR1-C3435T, CYP3A5-A6986G, CYP2C19-G681A, CYP2C19-G636A) from blood samples derived from liver transplant patients. The SNP detection system is comprised of three processes: multiplex PCR, single base extension with fluorescently labelled di-deoxy-nucleotides and detection by DNA microarray. The entire workflow of this system completes within 5 h. The final genotype call was obtained statistically by Mahalanobis distance which was calculated from the bi-coloured fluorescent signals detected by the microarray. In order to detect the six SNPs, this system required only 50 copies of genomic DNA, and the obtained detection calls completely matched with the results by the sequencing-based genotyping method. With the high sensitivity and rapid processing, our SNP detection system utilizing ultrasensitive microarray is a promising device applicable for diagnostic utility.
UR - http://www.scopus.com/inward/record.url?scp=78049475252&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=78049475252&partnerID=8YFLogxK
U2 - 10.1093/jb/mvq087
DO - 10.1093/jb/mvq087
M3 - Article
C2 - 20716514
AN - SCOPUS:78049475252
VL - 148
SP - 557
EP - 563
JO - Journal of Biochemistry
JF - Journal of Biochemistry
SN - 0021-924X
IS - 5
ER -