Center for Medical Genetics & Hunan Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410078, China.
Biosensors (Basel). 2021 May 14;11(5):154. doi: 10.3390/bios11050154.
Spinal muscular atrophy (SMA) is characterized by severe lethality and irreversible progression. Early diagnosis of SMA is of more practical significance with the emergence of effective therapy. However, existing techniques to identify SMA patients rely on cumbersome instruments, hindering their accessibility and application. An SMA-Cas12a-strip assay was developed with the integration of Cas12a-based nucleic acid detection, isothermal amplification, and lateral flow strip. The analytical performance of the assay was assessed with clinical samples. To explore its extensible utility, various specimens were tested. Validated with 168 clinical samples, the sensitivity and specificity of the SMA-Cas12a-strip assay were both 100%. The minimum detectable concentration of genomic DNA containing the target gene achieved 526 aM. The assay was compatible with specimens from several sources, and the turnaround time could be within 1.5 h. We developed a simple, cost-effective, and highly sensitive and specific assay to detect SMA patients. With little and field-portable equipment, the assay holds great promise in the detection of SMA patients, particularly in low-resource regions.
脊髓性肌萎缩症(SMA)的特点是致死率高且病情不可逆转。随着有效疗法的出现,早期诊断 SMA 更具实际意义。然而,目前用于识别 SMA 患者的技术依赖于繁琐的仪器,限制了其可及性和应用。本研究通过整合基于 Cas12a 的核酸检测、等温扩增和横向流动条带,开发了一种 SMA-Cas12a 检测条。采用临床样本评估该方法的分析性能,并通过测试各种标本来探索其可扩展性。经过 168 例临床样本验证,SMA-Cas12a 检测条的灵敏度和特异性均为 100%。最小可检测浓度为包含目标基因的基因组 DNA 为 526 aM。该检测条与多种来源的标本兼容,检测时间可在 1.5 小时内完成。我们开发了一种简单、经济高效、高灵敏度和特异性的检测方法,用于检测 SMA 患者。该检测条所需设备体积小且便于携带,有望用于 SMA 患者的检测,特别是在资源匮乏的地区。