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Rapid detection of apolipoprotein E genotypes in Alzheimer's disease using polymerase chain reaction-single strand conformation polymorphism.

Abstract

Apolipoprotein E (APOE) gene on chromosome 19q13.2 is encoded by three common alleles designated as epsilon2, epsilon3 and epsilon4. In Alzheimer's disease (AD) the epsilon4 allele is over-represented and is considered to be a major genetic risk factor. Several methods have been developed to determine APOE genotypes. Among them, polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) appears to be highly reliable. In this study, we improved the nonisotopic PCR-SSCP method for determining APOE genotypes in 42 cases of AD patients, 40 cases of non-AD dementia patients, and 49 cases of age-matched controls. DNA from the target sequence on APOE was amplified by PCR from peripheral blood genomic DNA. PCR products were electrophoresed in a non-denaturing polyacrylamide gel and visualized by silver staining. We found that the epsilon4 allele had a significantly high frequency of occurrence in AD patients (33.3%) compared with age-matched controls (13.3%) (chi(2) = 10.43, p = 0.001) and non-AD dementia (10%) (chi(2) = 13.02, p<0.001) whereas the epsilon3 allele was of high frequency in non-AD dementia (90%) compared with age-matched controls (85.7%) and AD patients (66.7%). APOE epsilon4 homozygotes were found only in AD groups. On the other hand, the epsilon2 allele was found only in an age-matched control. This study confirmed that the APOE psilon4 allele is a risk factor in Thai AD subjects and that the PCR-SSCP method is a rapid and useful means of detecting the APOE genotype in AD.

Authors+Show Affiliations

Neuro-Behavioural Biology Center, Institute of Science and Technology for Research and Development, Mahidol University, Nakhon Pathom, Bangkok.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

17121308

Citation

Kamruecha, Worawan, et al. "Rapid Detection of Apolipoprotein E Genotypes in Alzheimer's Disease Using Polymerase Chain Reaction-single Strand Conformation Polymorphism." The Southeast Asian Journal of Tropical Medicine and Public Health, vol. 37, no. 4, 2006, pp. 793-7.
Kamruecha W, Chansirikarnjana S, Nimkulrat E, et al. Rapid detection of apolipoprotein E genotypes in Alzheimer's disease using polymerase chain reaction-single strand conformation polymorphism. Southeast Asian J Trop Med Public Health. 2006;37(4):793-7.
Kamruecha, W., Chansirikarnjana, S., Nimkulrat, E., Udommongkol, C., Wongmek, W., & Thangnipon, W. (2006). Rapid detection of apolipoprotein E genotypes in Alzheimer's disease using polymerase chain reaction-single strand conformation polymorphism. The Southeast Asian Journal of Tropical Medicine and Public Health, 37(4), 793-7.
Kamruecha W, et al. Rapid Detection of Apolipoprotein E Genotypes in Alzheimer's Disease Using Polymerase Chain Reaction-single Strand Conformation Polymorphism. Southeast Asian J Trop Med Public Health. 2006;37(4):793-7. PubMed PMID: 17121308.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Rapid detection of apolipoprotein E genotypes in Alzheimer's disease using polymerase chain reaction-single strand conformation polymorphism. AU - Kamruecha,Worawan, AU - Chansirikarnjana,Sirintorn, AU - Nimkulrat,Ekapot, AU - Udommongkol,Chesda, AU - Wongmek,Wanna, AU - Thangnipon,Wipawan, PY - 2006/11/24/pubmed PY - 2007/2/28/medline PY - 2006/11/24/entrez SP - 793 EP - 7 JF - The Southeast Asian journal of tropical medicine and public health JO - Southeast Asian J Trop Med Public Health VL - 37 IS - 4 N2 - Apolipoprotein E (APOE) gene on chromosome 19q13.2 is encoded by three common alleles designated as epsilon2, epsilon3 and epsilon4. In Alzheimer's disease (AD) the epsilon4 allele is over-represented and is considered to be a major genetic risk factor. Several methods have been developed to determine APOE genotypes. Among them, polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) appears to be highly reliable. In this study, we improved the nonisotopic PCR-SSCP method for determining APOE genotypes in 42 cases of AD patients, 40 cases of non-AD dementia patients, and 49 cases of age-matched controls. DNA from the target sequence on APOE was amplified by PCR from peripheral blood genomic DNA. PCR products were electrophoresed in a non-denaturing polyacrylamide gel and visualized by silver staining. We found that the epsilon4 allele had a significantly high frequency of occurrence in AD patients (33.3%) compared with age-matched controls (13.3%) (chi(2) = 10.43, p = 0.001) and non-AD dementia (10%) (chi(2) = 13.02, p<0.001) whereas the epsilon3 allele was of high frequency in non-AD dementia (90%) compared with age-matched controls (85.7%) and AD patients (66.7%). APOE epsilon4 homozygotes were found only in AD groups. On the other hand, the epsilon2 allele was found only in an age-matched control. This study confirmed that the APOE psilon4 allele is a risk factor in Thai AD subjects and that the PCR-SSCP method is a rapid and useful means of detecting the APOE genotype in AD. SN - 0125-1562 UR - https://www.unboundmedicine.com/medline/citation/17121308/Rapid_detection_of_apolipoprotein_E_genotypes_in_Alzheimer's_disease_using_polymerase_chain_reaction_single_strand_conformation_polymorphism_ L2 - https://medlineplus.gov/alzheimersdisease.html DB - PRIME DP - Unbound Medicine ER -