Wijaya Yogik Onky Silvana, Ar Rochmah Mawaddah, Nurputra Dian Kesumapramudya, Farmawati Arta
Department of Biochemistry, Faculty of Medicine, Public Health, and Nursing, Universitas Gadjah Mada, Jalan Farmako, Yogyakarta, 55281, Indonesia.
Department of Neurology, Faculty of Medicine, Public Health, and Nursing, Universitas Gadjah Mada, Jalan Farmako, Yogyakarta, 55281, Indonesia.
BMC Biotechnol. 2025 Feb 14;25(1):17. doi: 10.1186/s12896-024-00938-2.
Spinal muscular atrophy (SMA) is a devastating neuromuscular condition resulting from the loss of the survival motor neuron 1 (SMN1) gene. Precise genetic testing has become essential after the authorization of several potent medications. To achieve this objective, the use of dried blood spot (DBS) has assured convenient and extensive testing from a distance. Nevertheless, developing countries such as Indonesia sometimes lack access to standard filter papers like FTA or Guthrie cards for DBS processing. Here, we aim to develop a cellulose-based card as an alternative filter paper for DBS preparation suitable for the genetic testing of SMA including but not limited to a direct polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and multiplex allele-specific amplification (multi-ASA).
An in-house paper was developed from a 180 gsm cellulose-based paper and was used for DBS preparation. The performance of dried blood spotted on the cellulose-based card (DBSc) was compared to pure genomic DNA (gDNA) isolate and dried blood spotted on FTA cards (DBSf) for genetic testing. The results of the genetic testing of our cellulose-based card were completely matched with those of gDNA and DBSf in both direct PCR-RFLP and Multi-ASA to separate SMN1 from SMN2. In addition, after three months of storing, the DBSc continued to exhibit a clear result, suggesting its high stability for DNA storage.
Our cellulose-based card has the potential to be used for DBS carrier and for further genetic testing using PCR. Our findings can assist physicians in sending DBS samples from SMA suspicion cases to genetic testing centers, thereby preventing diagnosis delay or misdiagnosis.
脊髓性肌萎缩症（SMA）是一种由生存运动神经元1（SMN1）基因缺失导致的严重神经肌肉疾病。在几种有效药物获批后，精确的基因检测变得至关重要。为实现这一目标，使用干血斑（DBS）确保了远距离的便捷和广泛检测。然而，像印度尼西亚这样的发展中国家有时缺乏用于DBS处理的标准滤纸，如FTA或Guthrie卡。在此，我们旨在开发一种基于纤维素的卡片作为DBS制备的替代滤纸，适用于SMA的基因检测，包括但不限于直接聚合酶链反应-限制性片段长度多态性（PCR-RFLP）和多重等位基因特异性扩增（multi-ASA）。
用180克重的纤维素基纸开发了一种自制纸张用于DBS制备。将基于纤维素的卡片（DBSc）上的干血斑性能与纯基因组DNA（gDNA）分离物以及FTA卡片（DBSf）上的干血斑进行基因检测比较。在直接PCR-RFLP和Multi-ASA中，我们基于纤维素的卡片的基因检测结果与gDNA和DBSf的结果完全匹配，以区分SMN1和SMN2。此外，储存三个月后，DBSc仍显示清晰结果，表明其对DNA储存具有高稳定性。
我们基于纤维素的卡片有潜力用作DBS载体并用于使用PCR的进一步基因检测。我们的发现可帮助医生将来自SMA疑似病例的DBS样本送往基因检测中心，从而防止诊断延迟或误诊。
