Guo Mei,Lei Jie,Yang Zhao,Chen Qinghai.Detection of CYP2C8 gene 139A/G single nucleotide polymorphism based on molecular beacon technology[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(11):1075-1079.
Guo Mei,Lei Jie,Yang Zhao,Chen Qinghai.Detection of CYP2C8 gene 139A/G single nucleotide polymorphism based on molecular beacon technology[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(11):1075-1079.DOI:
Objective To design a new fracture-connective hairpin probe based on the technique of molecular beacons
and then to establish and optimize a new method to detect the 139A/G single nucleotide polymorphism (SNP) of CYP2C8 gene by the probe. Methods Firstly
the fracture-connective hairpin probe was designed through the bioinformatic software
and then the desired probes and target sequences were synthesized. The optimal temperature and concentration ratio of probe were explored for the reaction system. At last
this method was performed to detect the 139A/G SNP of CYP2C8 gene. Results The optimal hybridization temperature for the fracture-connective hairpin probe was 40 ℃ to distinguish the SNP of target sequence
and the optimal hybridization ratio was 1∶4 of fluorescence probe to quenching probe. Under the condition of optimal temperature and ratio
the fluorescent intensity of the reaction system of the fracture-connective hairpin probe and CYP2C8 gene primary sequence was 7 632.0±8.7 RFU
while that of of the fracture-connective hairpin probe and CYP2C8 gene mutational sequence was 6 425.7±6.1 RFU. There was difference significance between the 2 reaction systems (P0.05). Conclusion Our new type of fracture-connective hairpin probe can effectively distinguish the primary and mutational sequences
and can be used to detect the 139A/G SNP of CYP2C8 gene. This study lays experimental foundation for the rapid and high-throughput SNP detection.