[Clinical application of real-time PCR for the detection of genetic mutations underlying spinal muscular atrophy].
OBJECTIVE
To 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.


METHODS
Thirty-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.


RESULTS
Genotyping 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.


CONCLUSIONS
The 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.
Molecular Diagnostics Laboratory, Xiamen Maternity and Child Health Care Hospital, Xiamen, Fujian 361003, P.R. China. zhou_yulin@126.com.
Yu
Y
Jiang
Guilan
G
Peng
Qichang
Q
Wu
Yulin
Y
Zhou
D018628:Gene Dosage; D006801:Humans; D009134:Muscular Atrophy, Spinal / Q000235:genetics*; D009154:Mutation; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D055533:Survival of Motor Neuron 1 Protein / Q000235:genetics
C526471:SMN1 protein, human; D055533:Survival of Motor Neuron 1 Protein