Evaluation and comparison of three assays for molecular detection of spinal muscular atrophy
| Autor: | Ti-Zhen Yan, Ze-Yan Zhong, Xu-Hui Tan, Wan-Jun Zhou, Jian Zeng, Xiang-Min Xu, Ping Fang, Liang Li, Jian-Sheng Xie |
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| Rok vydání: | 2017 |
| Předmět: |
Genetic Markers
0301 basic medicine Pathology medicine.medical_specialty DNA Copy Number Variations Genotype Clinical Biochemistry SMN1 Biology Muscular Atrophy Spinal 03 medical and health sciences 0302 clinical medicine Multiplex polymerase chain reaction medicine Humans Multiplex Multiplex ligation-dependent probe amplification Sequence Deletion Reproducibility Biochemistry (medical) Reproducibility of Results General Medicine SMA Survival of Motor Neuron 1 Protein Molecular biology Neuronal Apoptosis-Inhibitory Protein Survival of Motor Neuron 2 Protein 030104 developmental biology Real-time polymerase chain reaction Multiplex Polymerase Chain Reaction 030217 neurology & neurosurgery |
| Zdroj: | Clinical Chemistry and Laboratory Medicine (CCLM). 55 |
| ISSN: | 1437-4331 1434-6621 |
| DOI: | 10.1515/cclm-2016-0275 |
| Popis: | Background Spinal muscular atrophy (SMA) is mainly caused by deletions in SMA-related genes. The objective of this study was to develop gene-dosage assays for diagnosing SMA. Methods A multiplex, quantitative PCR assay and a CNVplex assay were developed for determining the copy number of SMN1, SMN2, and NAIP. Reproducibility and specificity of the two assays were compared to a multiple ligation-dependent probe amplification (MLPA) assay. To evaluate reproducibility, 30 samples were analyzed three times using the three assays. A total of 317 samples were used to assess the specificity of the two assays. Results The multiplex quantitative PCR (qPCR) assay had higher reproducibility. Intra-assay CVs were 3.01%-8.52% and inter-assay CVs were 4.12%-6.24%. The CNVplex assay had ratios that were closer to expected (0.49-0.5 for one copy, 1.03-1.0 for two copies, and 1.50-1.50 for three copies). Diagnostic accuracy rates for the two assays were 100%. Conclusions The multiplex qPCR assay was a simple, rapid, and cost-effective method for routine SMA diagnosis and carrier screening. The CNVplex assay could be used to detect SMAs with complicated gene structures. The assays were reliable and could be used as alternative methods for clinical diagnosis of SMA. |
| Databáze: | OpenAIRE |
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