关键词: 聚合酶链反应, 载脂蛋白A5, 2型糖尿病

Abstract: Objective　To establish fluorescence quantitation polymerase chain reaction (PCR) based on TaqMan-MGB probe for detecting apolipoprotein A5（apo A5）gene. Methods　The assay, which was based on specific primers and TaqMan-MGB probe from apo A5 gene, was performed to reconstruct DNA fragment of pPCR-Script-apoA5. The fluorescence quantitation PCR with TaqMan-MGB probe and the standard curve were established. The sensitivity， specificity and repeatability of the TaqMan-MGB probe fluorescence quantitation PCR were determined. Results　A fine linear relationship between the threshold cycle (Ct) values of the developed standard curve and the copy number of template was observed (r=0.998). The sensitivity of the assay was 10³ copies/μL, the amplification efficiency of assay was 110.2%, and the repeatability was good.Conclusions　The fluorescence quantitation PCR based on TaqMan-MGB probe for detecting the apo A5 gene is successfully developed with high sensitivity，specificity and repeatability.

Key words: Polymerase chain reaction, Apolipoprotein A5, Type 2 diabetes mellitus