MethyLight TaqMan® Assay for quantification of DNA methylation of Reelin (RELN) gene in schizophrenia: preliminary data

Introduction: Epigenetic changes, particularly DNA methylation are an important mechanism in the regulation of gene expression. Since DNA methylation occurs without genetic sequence alteration, it offers a new insight on the pathogenesis of various complex diseases like cancer and psychiatric diso...

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Main Authors:	Rahim, Mohd Nabil Fikri, Abdul Rahim, Nour El Huda, Mohd. Noor, Hanisah, Abdullah, Nor Zamzila, A.Talib, Norlelawati
Format:	Article
Language:	English
Published:	College of Pathologists, Academy of Medicine Malaysia 2015
Subjects:	R Medicine (General)
Online Access:	http://irep.iium.edu.my/49683/ http://irep.iium.edu.my/49683/ http://irep.iium.edu.my/49683/1/49683.pdf

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spelling	iium-496832018-05-21T02:18:42Z http://irep.iium.edu.my/49683/ MethyLight TaqMan® Assay for quantification of DNA methylation of Reelin (RELN) gene in schizophrenia: preliminary data Rahim, Mohd Nabil Fikri Abdul Rahim, Nour El Huda Mohd. Noor, Hanisah Abdullah, Nor Zamzila A.Talib, Norlelawati R Medicine (General) Introduction: Epigenetic changes, particularly DNA methylation are an important mechanism in the regulation of gene expression. Since DNA methylation occurs without genetic sequence alteration, it offers a new insight on the pathogenesis of various complex diseases like cancer and psychiatric disorder such as schizophrenia. Reelin (RELN) gene encodes an extracellular matrix protein that is responsible for neuronal migration during brain development. The methylation study of RELN is needed to clarify its association in schizophrenia. Materials and Methods: DNA samples were extracted from blood of patients diagnosed with schizophrenia in Hospital Tengku Ampuan Afzan, Pahang. The DNA samples were subjected to bisulfite conversion to enhance the methylation areas. Primers and probe were designed to cover CpG rich sites of RELN gene whilst ALU sequences were used as reference target. Sensitivity and specificity of the MethyLight TaqMan® assay were determined by running the assay in serial dilution for the distinction of methylated and unmethylated DNA. With the optimised assay, the DNA methylation of 10 schizophrenia patient samples was measured. Results: This study showed a decreasing detection of methylated alleles in each subsequent dilution using universal human methylated DNA with the Cq value for RELN gene increased from 16.89 to 29.68. We detected the percentage methylation ratio, which valued above zero for the RELN gene in all ten patients ranging from 0.91 to 1.17. Discussion: Our preliminary data suggests the presence of DNA methylation of RELN gene in schizophrenia patients. College of Pathologists, Academy of Medicine Malaysia 2015 Article PeerReviewed application/pdf en http://irep.iium.edu.my/49683/1/49683.pdf Rahim, Mohd Nabil Fikri and Abdul Rahim, Nour El Huda and Mohd. Noor, Hanisah and Abdullah, Nor Zamzila and A.Talib, Norlelawati (2015) MethyLight TaqMan® Assay for quantification of DNA methylation of Reelin (RELN) gene in schizophrenia: preliminary data. Malaysian Journal of Pathology, 37 (2). p. 191. ISSN 0126-8635 http://www.mjpath.org.my/2015/v37n2/
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English
topic	R Medicine (General)
spellingShingle	R Medicine (General) Rahim, Mohd Nabil Fikri Abdul Rahim, Nour El Huda Mohd. Noor, Hanisah Abdullah, Nor Zamzila A.Talib, Norlelawati MethyLight TaqMan® Assay for quantification of DNA methylation of Reelin (RELN) gene in schizophrenia: preliminary data
description	Introduction: Epigenetic changes, particularly DNA methylation are an important mechanism in the regulation of gene expression. Since DNA methylation occurs without genetic sequence alteration, it offers a new insight on the pathogenesis of various complex diseases like cancer and psychiatric disorder such as schizophrenia. Reelin (RELN) gene encodes an extracellular matrix protein that is responsible for neuronal migration during brain development. The methylation study of RELN is needed to clarify its association in schizophrenia. Materials and Methods: DNA samples were extracted from blood of patients diagnosed with schizophrenia in Hospital Tengku Ampuan Afzan, Pahang. The DNA samples were subjected to bisulfite conversion to enhance the methylation areas. Primers and probe were designed to cover CpG rich sites of RELN gene whilst ALU sequences were used as reference target. Sensitivity and specificity of the MethyLight TaqMan® assay were determined by running the assay in serial dilution for the distinction of methylated and unmethylated DNA. With the optimised assay, the DNA methylation of 10 schizophrenia patient samples was measured. Results: This study showed a decreasing detection of methylated alleles in each subsequent dilution using universal human methylated DNA with the Cq value for RELN gene increased from 16.89 to 29.68. We detected the percentage methylation ratio, which valued above zero for the RELN gene in all ten patients ranging from 0.91 to 1.17. Discussion: Our preliminary data suggests the presence of DNA methylation of RELN gene in schizophrenia patients.
format	Article
author	Rahim, Mohd Nabil Fikri Abdul Rahim, Nour El Huda Mohd. Noor, Hanisah Abdullah, Nor Zamzila A.Talib, Norlelawati
author_facet	Rahim, Mohd Nabil Fikri Abdul Rahim, Nour El Huda Mohd. Noor, Hanisah Abdullah, Nor Zamzila A.Talib, Norlelawati
author_sort	Rahim, Mohd Nabil Fikri
title	MethyLight TaqMan® Assay for quantification of DNA methylation of Reelin (RELN) gene in schizophrenia: preliminary data
title_short	MethyLight TaqMan® Assay for quantification of DNA methylation of Reelin (RELN) gene in schizophrenia: preliminary data
title_full	MethyLight TaqMan® Assay for quantification of DNA methylation of Reelin (RELN) gene in schizophrenia: preliminary data
title_fullStr	MethyLight TaqMan® Assay for quantification of DNA methylation of Reelin (RELN) gene in schizophrenia: preliminary data
title_full_unstemmed	MethyLight TaqMan® Assay for quantification of DNA methylation of Reelin (RELN) gene in schizophrenia: preliminary data
title_sort	methylight taqman® assay for quantification of dna methylation of reelin (reln) gene in schizophrenia: preliminary data
publisher	College of Pathologists, Academy of Medicine Malaysia
publishDate	2015
url	http://irep.iium.edu.my/49683/ http://irep.iium.edu.my/49683/ http://irep.iium.edu.my/49683/1/49683.pdf
first_indexed	2023-09-18T21:10:13Z
last_indexed	2023-09-18T21:10:13Z
_version_	1777411202853371904

MethyLight TaqMan® Assay for quantification of DNA methylation of Reelin (RELN) gene in schizophrenia: preliminary data

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