Establishment of a Nonradioactive Molecular Diagnosis of Fragile-X Syndrome

Zehra Oya UYGUNER

Makale detayları

Establishment of a Nonradioactive Molecular Diagnosis of Fragile-X Syndrome

Yazarlar:

Zehra Oya UYGUNER

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Medical Sciences 2000 Vol.30 Issue 3, pp.253-260

Belge Tipi:

Makale

Yayın Tarihi:

2/2000

Belge Dili:

Türkçe

Document ID (Org No.):

2000757628

Sıklık:

İki Aylık

eISSN:

1303-6165

pISSN:

1300-0144

Anahtar kelimeler:

Özet:

 

Fragile-X syndrome is a hereditary  dynamic mutation disorder, predominantly  caused by a large expansion of CGG  trinucleotide repeats in the FMR 1 gene  leading to methylation and down regulation  of transcription of the gene. For the  molecular diagnosis of the disease, the repeat  locus in FMR 1 gene is primarily detected by  Southern blotting with radioactively labeled  probes. Unusually GC rich composition of the  expanded region caused technical difficulties  during PCR based testing and therefore is not  performed by most diagnostic laboratories.  We established a Southern blot method and a  novel PCR protocol which enables the  amplification of normal, premutated and full  mutated alleles. In both techniques bands are  visualized by digoxigenin labeled probe and  chemiluminescent detection. Although  diagnosis of fragile-X was possible by  cytogenetic analysis in our division, it was not  possible to diagnose premutation carriers or  mosaic fragile-X patients. This newly  established nonradioactive PCR and Southern  blot analysis will provide routine detection of  full fragile-X mutations and premutations in  our laboratory and therefore enable us to  offer accurate genetic counseling and  prenatal diagnosis.

 

Fragile-X syndrome is a hereditary  dynamic mutation disorder, predominantly  caused by a large expansion of CGG  trinucleotide repeats in the FMR 1 gene  leading to methylation and down regulation  of transcription of the gene. For the  molecular diagnosis of the disease, the repeat  locus in FMR 1 gene is primarily detected by  Southern blotting with radioactively labeled  probes. Unusually GC rich composition of the  expanded region caused technical difficulties  during PCR based testing and therefore is not  performed by most diagnostic laboratories.  We established a Southern blot method and a  novel PCR protocol which enables the  amplification of normal, premutated and full  mutated alleles. In both techniques bands are  visualized by digoxigenin labeled probe and  chemiluminescent detection. Although  diagnosis of fragile-X was possible by  cytogenetic analysis in our division, it was not  possible to diagnose premutation carriers or  mosaic fragile-X patients. This newly  established nonradioactive PCR and Southern  blot analysis will provide routine detection of  full fragile-X mutations and premutations in  our laboratory and therefore enable us to  offer accurate genetic counseling and  prenatal diagnosis.

Makale detayları

Establishment of a Nonradioactive Molecular Diagnosis of Fragile-X Syndrome

Yazarlar:

Zehra Oya UYGUNER

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Medical Sciences 2000 Vol.30 Issue 3, pp.253-260

Belge Tipi:

Makale

Yayın Tarihi:

2/2000

Belge Dili:

Türkçe

Document ID (Org No.):

2000757628

Sıklık:

İki Aylık

eISSN:

1303-6165

pISSN:

1300-0144

Fragile-X

molecular diagnosis

nonradioactive labeling

polymerase chain reaction.

Sayı

Makale detayları

Establishment of a Nonradioactive Molecular Diagnosis of Fragile-X Syndrome

Yazarlar: Zehra Oya UYGUNER

Yayıncı: TUBITAK

Dergi: Turkish Journal of Medical Sciences 2000 Vol.30 Issue 3, pp.253-260

Belge Tipi: Makale

Yayın Tarihi: 2/2000

Belge Dili: Türkçe

Document ID (Org No.): 2000757628

Sıklık: İki Aylık

eISSN: 1303-6165

pISSN: 1300-0144

Fragile-X

molecular diagnosis

nonradioactive labeling

polymerase chain reaction.

Sayı

Yazarlar:

Zehra Oya UYGUNER

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Medical Sciences 2000 Vol.30 Issue 3, pp.253-260

Belge Tipi:

Makale

Yayın Tarihi:

2/2000

Belge Dili:

Türkçe

Document ID (Org No.):

2000757628

Sıklık:

İki Aylık

eISSN:

1303-6165

pISSN:

1300-0144