Department of Pediatrics, All India Institute of Medical Science Jodhpur, Jodhpur, Rajasthan, India.
Department of Medical Genetics, All India Institute of Medical Science Jodhpur, Jodhpur, Rajasthan, India.
Genet Test Mol Biomarkers. 2024 May;28(5):207-212. doi: 10.1089/gtmb.2023.0073. Epub 2024 Mar 27.
Spinal muscular atrophy (SMA) is an autosomal recessive neuromuscular life-threatening disorder. Owing to high carrier frequency, population-wide SMA screening to quantify the copy number of gene is recommended by American College of Medical Genetics and Genomics. An accurate, reliable, short runaround time and cost-effective method may be helpful in mass population screening for SMA. Multiplex ligation-dependent probe amplification (MLPA) is a gold standard to estimate the copy number variation (CNV) for and genes. In this study, we validated droplet digital polymerase chain reaction (ddPCR) for the determination of CNV for both and exon 7 for a diagnostic purpose. In total, 66 clinical samples were tested using ddPCR, and results were compared with the MLPA as a reference test. For all samples, CNV for and exon 7 was consentaneous between ddPCR and MLPA test results (κ = 1.000, < 0.0001). In addition, ddPCR also showed a significant acceptable degree of test repeatability, coefficient of variation < 4%. ddPCR is expected to be utilitarian for CNV detection for carrier screening and diagnosis of SMA. ddPCR test results for CNV detection for / exon 7 are concordant with the gold standard. ddPCR is a more cost-effective and time-saving diagnostic test for SMA than MLPA. Furthermore, it can be used for population-wide carrier screening for SMA.
脊髓性肌萎缩症（SMA）是一种常染色体隐性神经肌肉致死性疾病。由于高携带率，美国医学遗传学与基因组学学院建议对人群进行 SMA 基因拷贝数的全面筛查。一种准确、可靠、周转时间短且具有成本效益的方法可能有助于 SMA 的大规模人群筛查。多重连接依赖性探针扩增（MLPA）是评估 和 基因拷贝数变异（CNV）的金标准。在这项研究中，我们验证了用于确定 和 外显子 7 的 CNV 的数字微滴式 PCR（ddPCR），以便用于诊断目的。总共使用 ddPCR 测试了 66 个临床样本，结果与作为参考测试的 MLPA 进行了比较。对于所有样本，ddPCR 和 MLPA 测试结果的 和 外显子 7 的 CNV 一致（κ=1.000，<0.0001）。此外，ddPCR 还显示出具有显著可接受的测试重复性程度，变异系数<4%。ddPCR 有望用于携带者筛查和 SMA 诊断中的 CNV 检测。用于 / 外显子 7 的 CNV 检测的 ddPCR 测试结果与金标准一致。ddPCR 是一种比 MLPA 更具成本效益和更省时的 SMA 诊断测试。此外，它可用于 SMA 的人群筛查。
