All issues > Volume 48(10); 2005

Original Article

Korean J Pediatr. 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 JS Kang¹, Se Rim SR Kim², Sun Young SY Kim², Chan Uhng CU Joo¹, Soo Chul SC Cho¹, Pyoung Han PH 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

Correspondence Pyoung Han PH 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.

Keywords :Ghrelin , Polymorphism , PCR-RFLP , PCR-SSCP , ARMS