Angiotensin-converting enzyme insertion/deletion polymorphism genotyping error: the cause and a possible solution to the problem

被引:9
作者
Saracevic, Andrea [1 ]
Simundic, Ana-Maria [1 ]
Celap, Ivana [1 ]
Luzanic, Valentina [1 ]
机构
[1] Univ Hosp Ctr Sestre Milosrdnice, Univ Dept Chem, Zagreb 10000, Croatia
关键词
Angiotensin converting enzyme; Polymorphism; Genetic; Genotyping techniques; Polymerase chain reaction; GENE POLYMORPHISM; DELETION POLYMORPHISM; HETEROZYGOTES; AMPLIFICATION; FREQUENCY;
D O I
10.1007/s11033-013-2537-z
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Rigat and colleagues were the first ones to develop a rapid PCR-based assay for identifying the angiotensin converting enzyme insertion/deletion (I/D) polymorphism. Due to a big difference between the length of the wild-type and mute alleles the PCR method is prone to mistyping because of preferential amplification of the D allele causing depicting I/D heterozygotes as D/D homozygotes. The aim of this study was to investigate whether this preferential amplification can be repressed by amplifying a longer DNA fragment in a so called Long PCR protocol. We also aimed to compare the results of genotyping using five different PCR protocols and to estimate the mistyping rate. The study included 200 samples which were genotyped using standard method used in our laboratory, a stepdown PCR, PCR protocol with the inclusion of 4 % DMSO, PCR with the use of insertion specific primers and new Long PCR method. The results of this study have shown that accurate ACE I/D polymorphism genotyping can be accomplished with the standard and the Long PCR method. Also, as of our results, accurate ACE I/D polymorphism genotyping can be accomplished regardless of the method used. Therefore, if the standard method is optimized more cautiously, accurate results can be obtained by this simple, inexpensive and rapid PCR protocol.
引用
收藏
页码:4459 / 4463
页数:5
相关论文
共 18 条
[1]  
Chiang FT, 1998, CLIN CHEM, V44, P1353
[2]   The I/D polymorphism of the ACE1 gene is not associated with ischaemic stroke in Spanish individuals [J].
Domingues-Montanari, S. ;
Fernandez-Cadenas, I. ;
del Rio-Espinola, A. ;
Mendioroz, M. ;
Ribo, M. ;
Obach, V. ;
Marti-Fabregas, J. ;
Freijo, M. ;
Serena, J. ;
Corbeto, N. ;
Chacon, P. ;
Alvarez-Sabin, J. ;
Montaner, J. .
EUROPEAN JOURNAL OF NEUROLOGY, 2010, 17 (11) :1390-1392
[3]  
Faure-Delanef L, 1998, CLIN CHEM, V44, P2083
[4]   Simple detection of large InDeLS by DHPLC: The ACE gene as a model [J].
Koyama, Renata Guedes ;
Castro, Rosa M. R. P. S. ;
De Mello, Marco Tulio ;
Tufik, Sergio ;
Pedrazzoli, Mario .
JOURNAL OF BIOMEDICINE AND BIOTECHNOLOGY, 2008,
[5]   Angiotensin-converting enzyme gene deletion allele increases the risk of left ventricular hypertrophy: evidence from a meta-analysis [J].
Li, Xiaobo ;
Li, Yuqiong ;
Jia, Nan ;
Guo, Shujie ;
Chu, Shaoli ;
Niu, Wenquan .
MOLECULAR BIOLOGY REPORTS, 2012, 39 (12) :10063-10075
[6]   A PROSPECTIVE EVALUATION OF AN ANGIOTENSIN-CONVERTING-ENZYME GENE POLYMORPHISM AND THE RISK OF ISCHEMIC-HEART-DISEASE [J].
LINDPAINTNER, K ;
PFEFFER, MA ;
KREUTZ, R ;
STAMPFER, MJ ;
GRODSTEIN, F ;
LAMOTTE, F ;
BURING, J ;
HENNEKENS, CH .
NEW ENGLAND JOURNAL OF MEDICINE, 1995, 332 (11) :706-711
[7]  
Nawaz SK, 2009, BIOCHEM MEDICA, V19, P36
[8]   Mistyping frequency of the angiotensin-converting enzyme gene polymorphism and an improved method for its avoidance [J].
Odawara, M ;
Matsunuma, A ;
Yamashita, K .
HUMAN GENETICS, 1997, 100 (02) :163-166
[9]   PCR DETECTION OF THE INSERTION DELETION POLYMORPHISM OF THE HUMAN ANGIOTENSIN CONVERTING ENZYME GENE (DCP1) (DIPEPTIDYL CARBOXYPEPTIDASE-1) [J].
RIGAT, B ;
HUBERT, C ;
CORVOL, P ;
SOUBRIER, F .
NUCLEIC ACIDS RESEARCH, 1992, 20 (06) :1433-1433
[10]   AN INSERTION DELETION POLYMORPHISM IN THE ANGIOTENSIN I-CONVERTING ENZYME GENE ACCOUNTING FOR HALF THE VARIANCE OF SERUM ENZYME LEVELS [J].
RIGAT, B ;
HUBERT, C ;
ALHENCGELAS, F ;
CAMBIEN, F ;
CORVOL, P ;
SOUBRIER, F .
JOURNAL OF CLINICAL INVESTIGATION, 1990, 86 (04) :1343-1346