Detection of CYPIAl polymorphisms with a colorimetric method based on mismatch hybridization

被引:8
作者
Wu, XM [1 ]
Zhou, YK [1 ]
Xu, SQ [1 ]
机构
[1] Huazhong Univ Sci & Technol, Tongji Med Coll, Inst Environm Med, Wuhan 430030, Peoples R China
关键词
CYPIAl; genetic polymorphism; detection; mismatch hybridization; ELISA;
D O I
10.1016/S0009-8981(02)00130-4
中图分类号
R446 [实验室诊断]; R-33 [实验医学、医学实验];
学科分类号
1001 ;
摘要
Background: CYP1A1 polymorphisms, which have been reported to be associated with an elevated risk of lung cancer, are usually detected through conventional methods such as PCR-restriction fragment length polymorphism, allele-specific PCR and single-strand conformational polymorphism. Methods: An assay that makes use of differences in thermal stability between perfect match and non-perfect match hybrids has been developed. Two oligonucleotides probes for each CYP1A1 polymorphic site were designed and labeled with digoxigenin. After hybridization with amplified DNA fragment, the hybrids were detected with a colorimetric method and the genotype were identified by calculating the ratio of signals obtained with two probes. Results: The ratios for three genotypes obtained from 50 samples can be divided into three distinctive nonoverlapped groups when applying to m(1) and m(2) sites of CYP1A1 locus, which demonstrated the feasibility of this assay to detect CYP1A1 polymorphisms. Conclusion: Compared with other methods, this assay has lower cost, is fast, simple and is suitable for a screening test in routine laboratory. (C) 2002 Published by Elsevier Science B.V.
引用
收藏
页码:103 / 109
页数:7
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