ApoE genotyping by real-time fluorescence PCR
- VernacularTitle:应用实时荧光PCR分析apoE3、4等位基因
- Author:
Bingbing YAN
;
Tao YU
;
Huahong FUNG
;
Rui QIN
;
Shangyou YOU
- Publication Type:Journal Article
- Keywords:
apolipoprotein E;
single nucleotide polymorphism (SNP);
real-time fluorescence polymerase chain reaction (real-time fluorescence PCR)
- From:
Chinese Journal of Clinical Laboratory Science
2006;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a new effective method by using real-time polymerase chain reaction (PCR) to detect single nucleotide polymorphism (SNP) typing for rapid identifying apolipoprotein E alleles.Methods To determine alleles of human apolipoprotein E genetic polymorphism at Cys112Arg locus was detected by PCR melting curve analysis with fluorophore SYBR Green I. In order to increase the speciality of SNP assays, high fidelity Taq polymerase was used. The reliability of SNP typing was validated by comparison with the results of direct DNA sequencing.Results Each sample was determined by double tubes, and two melting curves were analysis. As compared the Tm value of samples with the Tm of standard substance, the apoE genotype of samples was determined. The apoE genotype of 30 samples were E3/3 (27/30) and E3/4 (3/30) respectively, which was accordant with the results of PCR-RFLP and DNA sequencing.Conclusion The presented allelic assay was specific, easy to operate and applicable for discrimination of apolipoprotein E genotyping of human blood.
