Exploration Of Real Time PCR As A New Diagnostic Method Of Fragile X Syndrome

Fragile X Syndrome (FXS) is the most common form of inherited mental retardation in males caused by FMR1 gene abnormality associated with CGG repeats expansion and hypermethylation status of its promoter. Methylated alleles usually lead to transcriptional inhibition and consequently loss of Fragi...

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Main Author:	Elias, Marjanu Hikmah
Format:	Thesis
Language:	English
Published:	2010
Subjects:	R5-130.5 General works
Online Access:	http://eprints.usm.my/42483/1/MARJANU_HIKMAH_BINTI_ELIAS_HJ.pdf
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id	my-usm-ep.42483
record_format	uketd_dc
spelling	my-usm-ep.424832019-04-12T05:26:58Z Exploration Of Real Time PCR As A New Diagnostic Method Of Fragile X Syndrome 2010 Elias, Marjanu Hikmah R5-130.5 General works Fragile X Syndrome (FXS) is the most common form of inherited mental retardation in males caused by FMR1 gene abnormality associated with CGG repeats expansion and hypermethylation status of its promoter. Methylated alleles usually lead to transcriptional inhibition and consequently loss of Fragile X Mental Retardation Protein (FMRP) production. Chemical modification of cytosine to uracil by sodium bisulfite treatment has provided an additional method for the laboratory diagnosis of FXS, thus avoiding the use of the laborious Southern blot analysis, which is the gold standard test for FXS diagnosis. Thus, a study was done to explore a rapid, easy, reliable and cheap method for FXS diagnosis that can replace the laborious, time consuming and expensive Southern blot method. Genomic DNA was extracted from peripheral blood of 45 clinically diagnosed FXS patients. Samples were treated with sodium bisulfite followed by amplification using real-time multiplex methylation specific PCR (RT-M-MSPCR) with XIST gene as an internal control, followed by melting curve analysis. Our results showed all control samples with known methylation status were correctly diagnosed using Real Time Multiplex Methylation Specific PCR. For method validation purpose, the methylation status of other 45 male patients sample were also successfully diagnosed using our RT-M-MSPCR method and were concordance with the results of the Southern blot. 2010 Thesis http://eprints.usm.my/42483/ http://eprints.usm.my/42483/1/MARJANU_HIKMAH_BINTI_ELIAS_HJ.pdf application/pdf en public masters Universiti Sains Malaysia Pusat Pengajian Sains Perubatan
institution	Universiti Sains Malaysia
collection	USM Institutional Repository
language	English
topic	R5-130.5 General works
spellingShingle	R5-130.5 General works Elias, Marjanu Hikmah Exploration Of Real Time PCR As A New Diagnostic Method Of Fragile X Syndrome
description	Fragile X Syndrome (FXS) is the most common form of inherited mental retardation in males caused by FMR1 gene abnormality associated with CGG repeats expansion and hypermethylation status of its promoter. Methylated alleles usually lead to transcriptional inhibition and consequently loss of Fragile X Mental Retardation Protein (FMRP) production. Chemical modification of cytosine to uracil by sodium bisulfite treatment has provided an additional method for the laboratory diagnosis of FXS, thus avoiding the use of the laborious Southern blot analysis, which is the gold standard test for FXS diagnosis. Thus, a study was done to explore a rapid, easy, reliable and cheap method for FXS diagnosis that can replace the laborious, time consuming and expensive Southern blot method. Genomic DNA was extracted from peripheral blood of 45 clinically diagnosed FXS patients. Samples were treated with sodium bisulfite followed by amplification using real-time multiplex methylation specific PCR (RT-M-MSPCR) with XIST gene as an internal control, followed by melting curve analysis. Our results showed all control samples with known methylation status were correctly diagnosed using Real Time Multiplex Methylation Specific PCR. For method validation purpose, the methylation status of other 45 male patients sample were also successfully diagnosed using our RT-M-MSPCR method and were concordance with the results of the Southern blot.
format	Thesis
qualification_level	Master's degree
author	Elias, Marjanu Hikmah
author_facet	Elias, Marjanu Hikmah
author_sort	Elias, Marjanu Hikmah
title	Exploration Of Real Time PCR As A New Diagnostic Method Of Fragile X Syndrome
title_short	Exploration Of Real Time PCR As A New Diagnostic Method Of Fragile X Syndrome
title_full	Exploration Of Real Time PCR As A New Diagnostic Method Of Fragile X Syndrome
title_fullStr	Exploration Of Real Time PCR As A New Diagnostic Method Of Fragile X Syndrome
title_full_unstemmed	Exploration Of Real Time PCR As A New Diagnostic Method Of Fragile X Syndrome
title_sort	exploration of real time pcr as a new diagnostic method of fragile x syndrome
granting_institution	Universiti Sains Malaysia
granting_department	Pusat Pengajian Sains Perubatan
publishDate	2010
url	http://eprints.usm.my/42483/1/MARJANU_HIKMAH_BINTI_ELIAS_HJ.pdf
_version_	1747821074349817856

Exploration Of Real Time PCR As A New Diagnostic Method Of Fragile X Syndrome

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