Clinical application of real-time PCR for the detection of genetic mutations underlying spinal muscular atrophy.
- Author:
Yu JIANG
1
;
Guilan PENG
;
Qichang WU
;
Yulin ZHOU
Author Information
- Publication Type:Journal Article
- MeSH: Gene Dosage; Humans; Muscular Atrophy, Spinal; genetics; Mutation; Real-Time Polymerase Chain Reaction; methods; Survival of Motor Neuron 1 Protein; genetics
- From: Chinese Journal of Medical Genetics 2014;31(2):180-184
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo verify the reliability of real-time PCR for the detection of genetic mutations underlying spinal muscular atrophy (SMA) and establish quality control for clinical testing.
METHODSThirty-five patients, 61 first-degree relatives, 61 healthy controls and 7 prenatal cases which were previously genotyped by multiplex ligation-dependent probe amplification (MLPA) were tested with Roche LightCycler 480 and Bio-Rad CFX96 (TM) real-time PCR machines for relative quantification of copy number of SMN1 exon 7.
RESULTSGenotyping detected by relative quantitative real-time PCR were consistent with the results of MLPA. Both types of real-time PCR machines could accurately distinguish different SMN1 copy numbers despite certain systematic differences between the two platforms.
CONCLUSIONThe reliability of real-time PCR assay for detecting SMA depends on quality control. Standard database generated with known SMN1 copy number variations should be established for different instruments.
