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ISSN: 2574 -1241
ISSN: 2574 -1241
Received: August 9, 2018; Published: August 17, 2018
DOI: 10.26717/BJSTR.2018.08.001604
Introduction: Spinal muscular atrophy (SMA) is a severe neurodegenerative autosomal recessive disorder. Most of patients are caused by the homozygous absence of a region of exon 7 and exon 8 of the telomeric copy of the SMN gene on chromosome 5. Seting up a molecular diagnostic protocol for detecting SMNt mutation in single cell is basic to Preimplantation Gentic Diagnosis.
Patients and Methods: We test 4 patients and their parent. Lymphocytes of patients and their parent was isolated from fresh blood by ficoll. Taking a lymphocyte by stereoscopic microscope, lysiced the cell, amplifying exon 7 and exon 8 of SMNt gene by using a nested polymerase chain reaction, followed by DraI and DdeI restriction digest of the PCR enabling the important SMNt gene to be distinguished from the centromic SMNc gene which has no clinical phenotype to detect mutation. Electrophoresis PCR products after digesting by restriction enzyme and analysis.
Result: Four patients showed deletion in exon 7, exon 8 SMNt gene. This result is similar with the gene diagnosis from fresh blood.
Conclusion: We have successfully applied the technique of nested-PCR for the gene diagnosis of spinal muscular atrophy from single cell.
Keywords: Spinal Muscular Atrophy; SMN Gene; Nested PCR; Single Cell
Abstract | Introduction | Materials and Methods | Results and Discussion | Conclusion and Recommendations | References |
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