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Comparision of PCR-RFLP, PCR-SSCP, Amplication Refractory Mutation System(ARMS) in Leu72Met Polymorphism of Ghrelin Gene

Korean Journal of Pediatrics 2005;48(10):1068-1075.
Published online October 15, 2005.
Comparision of PCR-RFLP, PCR-SSCP, Amplication Refractory Mutation System(ARMS) in Leu72Met Polymorphism of Ghrelin Gene
Ju Sung Kang1, Se Rim Kim2, Sun Young Kim2, Chan Uhng Joo1, Soo Chul Cho1, Pyoung Han Hwang2
1Department of Pediatrics, Schoool of Medicine, Chonbuk National University, Jeonju, Korea
2Department of Research Institute of Clinical Medicine, Schoool of Medicine, Chonbuk National University, Jeonju, Korea
Ghrelin 유전자의 Leu72Met 다형성 분석에서 PCR-RFLP, PCR-SSCP, Amplication Refractory Mutation System(ARMS)의 비교분석
강주성1, 김세림2, 김선영2, 주찬웅1, 조수철1, 황평한2
1전북대학교 의과대학 소아과학교실
2전북대학교 의과대학 임상의학연구소
Correspondence: 
Pyoung Han Hwang, Email: hwaph@chonbuk.ac.kr
Abstract
Purpose
: The role of ghrelin, which promotes the secretion of growth hormone, was not well known until now. Recently it was found that the mutation of ghrelin gene is related to obesity and diabetes. This study is to find the screening method that can easily and effectively detect the polymorphism of Leu72Met in ghrelin gene of obesity patients and apply it to clinical usage.
Methods
: We compared PCR-RFLP, PCR-SSCP and ARMS methodologies for analyzing of the polymorphism of Leu72Met in ghrelin gene of obesity children, and also studied the merits and demerits of these methodologies.
Results
: In this study, we were able to find out the band of peculiar allele of Leu72Met in ghrelin gene using PCR-RFLP, PCR-SSCP and ARMS analyses. The polymorphism of Leu72Met in ghrelin gene determined by all above methodologies was in complete agreement. Compared to the PCR- RFLP and PCR-SSCP, ARMS analysis is simple, inexpensive and also consume less time. It is very sensitive to analyze the polymorphism and easy to understand the results of test.
Conclusion
: Though PCR-RFLP, PCR-SSCP and ARMS analyses were sensitive to analyze the polymorphism of Leu72Met in ghrelin gene, ARMS analysis appears to be more efficient than PCR-RFLP and PCR-SSCP. Therefore, we conclude that ARMS analysis is suitable to analyze the polymorphism of Leu72Met in ghrelin gene for large quantity of specimens.
Key Words: Ghrelin , Polymorphism , PCR-RFLP , PCR-SSCP , ARMS


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