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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| Format: | Thesis |
| Language: | English |
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2010
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| Online Access: | http://eprints.usm.my/42483/ http://eprints.usm.my/42483/1/MARJANU_HIKMAH_BINTI_ELIAS_HJ.pdf |
| _version_ | 1848879572645314560 |
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| author | Elias, Marjanu Hikmah |
| author_facet | Elias, Marjanu Hikmah |
| author_sort | Elias, Marjanu Hikmah |
| building | USM Institutional Repository |
| collection | Online Access |
| 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. |
| first_indexed | 2025-11-15T17:49:22Z |
| format | Thesis |
| id | usm-42483 |
| institution | Universiti Sains Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T17:49:22Z |
| publishDate | 2010 |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | usm-424832019-04-12T05:26:58Z http://eprints.usm.my/42483/ Exploration Of Real Time PCR As A New Diagnostic Method Of Fragile X Syndrome 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 NonPeerReviewed application/pdf en http://eprints.usm.my/42483/1/MARJANU_HIKMAH_BINTI_ELIAS_HJ.pdf Elias, Marjanu Hikmah (2010) Exploration Of Real Time PCR As A New Diagnostic Method Of Fragile X Syndrome. Masters thesis, Universiti Sains Malaysia. |
| spellingShingle | R5-130.5 General works Elias, Marjanu Hikmah Exploration Of Real Time PCR As A New Diagnostic Method Of Fragile X Syndrome |
| title | 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_short | 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 |
| topic | R5-130.5 General works |
| url | http://eprints.usm.my/42483/ http://eprints.usm.my/42483/1/MARJANU_HIKMAH_BINTI_ELIAS_HJ.pdf |