Marzuki, Marini
(2012)
Development Of EZ DNA Diagnostic Kit For The Detection Of Homozygous Deletion Of SMN1 Gene In Spinal Muscular Atrophy (SMA) Patients.
Masters thesis, Universiti Sains Malaysia.
Abstract
Spinal Muscular Atrophy (SMA) is the second most frequent fatal autosomal recessive
disorder of childhood. The incidence of this disease is approximately 1 in 10000 live
births. SMA is characterized by progressive muscle weakness resulting from
degeneration and loss of motor neurons in the anterior horn of spinal cord. The
responsible genes for SMA are Survival of Motor Neuron (SMN). SMN1 and SMN2
genes share over 99.8% sequence homology and they can be distinguished by base
changes in both exons 7 and 8. SMN1 gene is not detectable in majority of SMA cases
due to either deletion or conversion. Conventionally, the homozygous deletion of SMN1
gene is detected using Polymerase Chain Reaction-Restriction Enzyme (PCR-RE)
method. This method is time consuming, expensive and requires restriction enzyme
digestion and a considerably high amount of amplified DNA to be visible after
digestion. This may lead to false-negative results. To avoid these problems, we have
developed an alternative method using an allele-specific primer for the molecular
diagnosis of SMA which is more time-saving and cost-effective. The freeze dry platform
was applied to the multiplex AS-PCR for the development of a thermostabilize
diagnostic kit. A total of one hundred and forty three samples of clinically suspected
SMA were included in this study. Conventional molecular diagnosis using PCR-RE was
done to detect the presence or absent of SMN1 deletion in these samples.
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