›› 2012, Vol. 27 ›› Issue (5): 400-403.

Previous Articles     Next Articles

Development and evaluation of Real-time PCR and TaqMan-MGB Probe assay for detecting of apolipoprotein A5 gene

  

  1. 1.Medical College of Southeast University, Jiangsu Nanjing 210008, China;2.Department of Clinical Laboratory, Drum Tower Hospital Affiliated to Medical School of Nanjing University, Jiangsu Nanjing 210008, China
  • Received:2012-03-19 Online:2012-05-30 Published:2012-05-10

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