Ignite Creation Date:
2024-05-05 @ 11:51 AM
Last Modification Date:
2024-10-26 @ 9:15 AM
Study NCT ID:
NCT00155168
Status:
UNKNOWN
Last Update Posted:
2005-09-12
First Post:
2005-09-09
Brief Title:
Quantitative Analysis of SMN1 and SMN2 Gene Based on DHPLC System
Sponsor:
National Taiwan University Hospital
Organization:
National Taiwan University Hospital
Study Overview
Official Title:
Quantitative Analysis of SMN1 and SMN2 Gene Based on DHPLC System Establishing a Novel Highly Efficient and Reliable SMA Carrier Screening Test
Status:
UNKNOWN
Status Verified Date:
2004-04
Last Known Status:
RECRUITING
Delayed Posting:
No
If Stopped, Why?:
Not Stopped
Has Expanded Access:
False
If Expanded Access, NCT#:
N/A
Has Expanded Access, NCT# Status:
N/A
Acronym:
None
Brief Summary:
In this project we will establish the efficient and accurate gene dose determination system by combining the heterodulex analysis and gene dose analysis on DHPLC platform based on various quantitative and multiplex PCR strategies and applying on detecting the carriers- in- risk and patients with spinal muscular atrophyThis method is therefore based on the observation that the amount of PCR product generated from each site of amplification is proportional to the amount of starting template Detection of PCR products is carried out on DHPLC which provide the sensitivity required for the detection of the single-copy dosage changes
Detailed Description:
Proximal spinal muscular atrophy is an autosomal recessive disorder with an overall incidence of 1 in 10000 live births and a carrier frequency of 1 in 50 This severe neuromuscular disease is characterized by a degeneration and loss of alpha motor neurons of the spinal cord anterior horn cells which results in progressive symmetrical weakness atrophy of the proximal voluntary muscles and infant death
Two most identical copies of SMN gene telomeric SMN SMN1 and centromeric SMN SMN2 have been identified These two SMN genes are highly homologous and differing in only five nucleotide exchanges within their 3 regions These variations do not alter the encoded amino acids These nucleotide differences located in exons 7 and 8 allow SMN1 gene to be distinguished from SMN2 gene
It has been reported that more than 95 of SMA patients were homozygous deletion of SMN1 gene Therefore the detection of the absence of SMN1 can be a useful tool for SMA diagnosis Furthermore because of the high incidence of SMA the high carrier frequency of at least 1 in 50 the severity of the disease in the patients and the lack of effective of treatment carrier testing for SMN1 deletion is an important question in genetic counseling However a highly homologous SMN2 gene also exits and hampers the detection of the loss of SMN1 which makes the detection of SMA carrier test difficult
Here we present a new rapid simple and high reliable method to detect the SMN1 deletion and to determine the copy number of the SMN1 and SMN2 by denaturing high-performance liquid chromatography DHPLC DHPLC is a novel simple fast and non gel-base method that is very sensitive and specific for detection DNA variations Furthermore we describe a multiplex PCR strategy to determine the SMN1 and SMN2 genes copy number in order to avoid bias due to fluctuations in the copy number of SMN genes The assay uses the X-linked CYBB gene and KRIT1 gene as standards to determine the relative gene dosage of SMN1 and SMN2 genes We demonstrate that this assay is able to accurately distinguish 2 gene copies from 4 gene copies and it can identify SMA carriers and normal populations by the accurate determination of SMN copy number
This project will contribute to apply this novel fast and reliable tool for diagnosis of SMA and carrier detection of SMN1 and SMN2 genes by using DHPLC system
Two most identical copies of SMN gene telomeric SMN SMN1 and centromeric SMN SMN2 have been identified These two SMN genes are highly homologous and differing in only five nucleotide exchanges within their 3 regions These variations do not alter the encoded amino acids These nucleotide differences located in exons 7 and 8 allow SMN1 gene to be distinguished from SMN2 gene
It has been reported that more than 95 of SMA patients were homozygous deletion of SMN1 gene Therefore the detection of the absence of SMN1 can be a useful tool for SMA diagnosis Furthermore because of the high incidence of SMA the high carrier frequency of at least 1 in 50 the severity of the disease in the patients and the lack of effective of treatment carrier testing for SMN1 deletion is an important question in genetic counseling However a highly homologous SMN2 gene also exits and hampers the detection of the loss of SMN1 which makes the detection of SMA carrier test difficult
Here we present a new rapid simple and high reliable method to detect the SMN1 deletion and to determine the copy number of the SMN1 and SMN2 by denaturing high-performance liquid chromatography DHPLC DHPLC is a novel simple fast and non gel-base method that is very sensitive and specific for detection DNA variations Furthermore we describe a multiplex PCR strategy to determine the SMN1 and SMN2 genes copy number in order to avoid bias due to fluctuations in the copy number of SMN genes The assay uses the X-linked CYBB gene and KRIT1 gene as standards to determine the relative gene dosage of SMN1 and SMN2 genes We demonstrate that this assay is able to accurately distinguish 2 gene copies from 4 gene copies and it can identify SMA carriers and normal populations by the accurate determination of SMN copy number
This project will contribute to apply this novel fast and reliable tool for diagnosis of SMA and carrier detection of SMN1 and SMN2 genes by using DHPLC system
Study Oversight
Has Oversight DMC:
None
Is a FDA Regulated Drug?:
None
Is a FDA Regulated Device?:
None
Is an Unapproved Device?:
None
Is a PPSD?:
None
Is a US Export?:
None
Is an FDA AA801 Violation?:
None