title: A rapid and cost-effective method for genotyping apolipoprotein E gene polymorphism.
abstract: 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Fujian Provincial Key Laboratory of Neurodegenerative Disease and Aging Research, Institute of Neuroscience, Medical College, Xiamen University, Xiamen, 361102, China. zhongli1103@163.com.
authors.forename: Li
authors.initials: L
authors.lastname: Zhong
authors.affiliation: Fujian Provincial Key Laboratory of Neurodegenerative Disease and Aging Research, Institute of Neuroscience, Medical College, Xiamen University, Xiamen, 361102, China. xyzh1984@163.com.
authors.forename: Yong-Zhuang
authors.initials: YZ
authors.lastname: Xie
authors.affiliation: Fujian Provincial Key Laboratory of Neurodegenerative Disease and Aging Research, Institute of Neuroscience, Medical College, Xiamen University, Xiamen, 361102, China. caott@memorigen.com.
authors.forename: Tian-Tian
authors.initials: TT
authors.lastname: Cao
authors.affiliation: Fujian Provincial Key Laboratory of Neurodegenerative Disease and Aging Research, Institute of Neuroscience, Medical College, Xiamen University, Xiamen, 361102, China. wangzongqionline@126.com.
authors.forename: Zongqi
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Fujian Provincial Key Laboratory of Neurodegenerative Disease and Aging Research, Institute of Neuroscience, Medical College, Xiamen University, Xiamen, 361102, China. wtt987654321wtt@163.com.
authors.forename: Tingting
authors.initials: T
authors.lastname: Wang
authors.affiliation: Fujian Provincial Key Laboratory of Neurodegenerative Disease and Aging Research, Institute of Neuroscience, Medical College, Xiamen University, Xiamen, 361102, China. lixinxiu40@aliyun.com.
authors.forename: Xinxiu
authors.initials: X
authors.lastname: Li
authors.affiliation: Fujian Institute of Subtropical Botany, Xiamen, 361006, China. xmsrc@163.com.
authors.forename: Rui-Chi
authors.initials: RC
authors.lastname: Shen
authors.affiliation: Fujian Provincial Key Laboratory of Neurodegenerative Disease and Aging Research, Institute of Neuroscience, Medical College, Xiamen University, Xiamen, 361102, China. hxxu@xmu.edu.cn.
authors.forename: Huaxi
authors.initials: H
authors.lastname: Xu
authors.affiliation: Fujian Provincial Key Laboratory of Neurodegenerative Disease and Aging Research, Institute of Neuroscience, Medical College, Xiamen University, Xiamen, 361102, China. bu.guojun@mayo.edu.|Department of Neuroscience, Mayo Clinic, Jacksonville, FL, 32224, USA. bu.guojun@mayo.edu.
authors.forename: Guojun
authors.initials: G
authors.lastname: Bu
authors.affiliation: Fujian Provincial Key Laboratory of Neurodegenerative Disease and Aging Research, Institute of Neuroscience, Medical College, Xiamen University, Xiamen, 361102, China. chenxf@xmu.edu.cn.
authors.forename: Xiao-Fen
authors.initials: XF
authors.lastname: Chen
mesh_terms: D000483:Alleles; D001057:Apolipoproteins E / Q000235:genetics*; D005787:Gene Frequency / Q000235:genetics*; D020022:Genetic Predisposition to Disease; D005820:Genetic Testing / Q000191:economics / Q000379:methods; D005838:Genotype; D006801:Humans; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D012307:Risk Factors; D013997:Time Factors
chemical_list: D001057:Apolipoproteins E