A rapid and cost-effective method for genotyping apolipoprotein E gene polymorphism

Molecular Neurodegeneration

Xiao-Fen Chen³  Guojun Bu¹  Huaxi Xu³  Rui-Chi Shen²  Xinxiu Li³  Tingting Wang³  Zongqi Wang³  Tian-Tian Cao³  Yong-Zhuang Xie³  Li Zhong³ 

[1] Department of Neuroscience, Mayo Clinic, Jacksonville 32224, FL, USA;Fujian Institute of Subtropical Botany, Xiamen 361006, China;Fujian Provincial Key Laboratory of Neurodegenerative Disease and Aging Research, Institute of Neuroscience, Medical College, Xiamen University, Xiamen 361102, China

关键词: Polymorphism;    TaqMan;    Sequencing;    Real Time PCR;    APOE genotype;    Apolipoprotein E;   

Others  :  1235472
DOI  :  10.1186/s13024-016-0069-4

 received in 2015-11-30, accepted in 2016-01-04,  发布年份 2016

【 摘 要 】

Background

Apolipoprotein E (ApoE) is a major cholesterol carrier and plays an important role in maintaining lipid homeostasis both in the periphery and brain. Human APOE gene is polymorphic at two single nucleotides (rs429358 and rs7412) resulting in three different alleles (ε2, ε3 and ε4). ApoE isoforms modulate the risk for a variety of vascular and neurodegenerative diseases; thus, APOE genotyping is crucial for predicting disease risk and designing individualized therapy based on APOE genotype.

Results

We have developed an APOE genotyping method that is based on allele-specific PCR methodology adapted to Real Time PCR monitored by TaqMan probe. Rather than using TaqMan probes specific for the two polymorphic sites, only one TaqMan probe is used as the polymorphic alleles are recognized by site-specific PCR primers. Each genotyping assay can be completed within 90 minutes and is applicable to high-throughput analysis. Using this protocol, we genotyped a total of 1158 human DNA samples and obtained a 100 % concordance with the APOE genotype determined by sequencing analysis.

Conclusion

The APOE genotyping assay we have developed is accurate and cost-effective. In addition, our assay can readily be applied to genotyping large sample numbers. Therefore, our APOE genotyping method can be used for assessing the risk for a variety of vascular and neurodegenerative diseases that have been reported to be associated with APOE polymorphism.

【 授权许可】

   
2016 Zhong et al.

【 图 表 】

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