Spinal Muscular Atrophy Patient Detection and Carrier Screening Using Dried Blood Spots on Filter Paper
Spinal Muscular Atrophy Patient Detection and Carrier Screening Using Dried Blood Spots on Filter Paper
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DOI：
10.1089/gtmb.2011.0109
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发表时间：
2012-02-01
影响因子：
1.4
通讯作者：
Nishio, Hisahide
中科院分区：
文献类型：
--
作者：
Harahap, Nur Imma Fatimah;Harahap, Indra Sari Kusuma;Nishio, Hisahide
目的：脊髓性肌萎缩症是一种常见的常染色体隐性遗传性神经肌肉疾病。它是由SMN1的突变引起的，其临床严重性由SMN1的拷贝数变化所改变。根据以前的研究，SMN1外显子7的缺失在SMA患者中最常见。因此，利用这种遗传损伤的分子分析可能有助于SMA的诊断。不幸的是，在许多地理区域，医生没有最新的分子筛查技术可供立即使用。因此，为了解决这一问题，我们开发了一种利用滤纸上的干血斑(DBS)进行SMA诊断的系统，以便于远程诊断。方法：在本研究中，我们验证了Flinders Technology Associates(FTA)滤纸上的DBS在SMN1外显子7缺失和SMN1和SMN2拷贝数变异检测中的适用性。为了检测SMN1基因外显子7的缺失，用FTA滤纸上的FTA滤纸提取的DNA进行聚合酶链式反应-限制性片段长度多态性分析。为了确定SMN1和SMN2的拷贝数，我们用相同的血液样本进行了SYBR Green实时荧光定量聚合酶链式反应。结果：PTA滤纸DBS检测结果与新鲜血液标本检测结果完全一致。这表明滤纸上的DBS是一种可靠的SMA患者检测和携带者筛查方法。结论：SMA诊断系统与滤纸上DBS的邮寄相结合，将有利于在缺乏或无法获得具有分子功能的诊断设施的地区的神经肌肉疾病患者。
Aim: Spinal muscular atrophy (SMA) is a common autosomal recessive neuromuscular disorder. It is caused by mutations in the SMN1, and its clinical severity is modified by copy number variations of the SMN2. According to previous studies, deletion of SMN1 exon 7 is the most frequently observed in patients with SMA. Therefore, molecular analyses exploiting this genetic lesion could be beneficial in the diagnosis of SMA. Unfortunately, in many geographical regions, physicians do not have the latest molecular screening technologies at their immediate disposal. Thus, to overcome this issue, we developed an SMA-diagnosing system using dried blood spots (DBS) placed on filter paper to facilitate remote diagnosis. Methods: In this study, we validate the applicability of DBS on Flinders Technology Associates (FTA) filter paper for detecting SMN1 exon 7 deletions and copy number variations of SMN1 and SMN2. To detect exon 7 deletions in SMN1, polymerase chain reaction (PCR)-restriction fragment length polymorphism analysis was conducted by using DNA extracted from the CBS on FTA filter paper that had been stored at room temperature for a period of up to 4 years. To determine the copy numbers of SMN1 and SMN2, we carried out SYBR green-based real-time PCR by using the same blood specimens. Results: The results obtained from the DBS on PTA filter paper were in complete concordance with those analyses using fresh blood specimens. This indicates that DBS on filter papers is a reliable method for SMA patient detection and carrier screenings. Conclusion: The SMA-diagnosing system, combined with the mailing of DBS on filter paper, will be beneficial for patients suffering from neuromuscular disorders in areas with limited or no access to diagnostic facilities with molecular capabilities.