Rapid diagnosis of spinal muscular atrophy using high-resolution melting analysis.
Rapid diagnosis of spinal muscular atrophy using high-resolution melting analysis.
复制标题
使用高分辨率熔解分析快速诊断脊髓性肌萎缩症
DOI：
10.1186/1471-2350-10-45
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发表时间：
2009-05-29
影响因子：
--
通讯作者：
Wang N
中科院分区：
文献类型：
--
作者：
Chen WJ;Dong WJ;Lin XZ;Lin MT;Murong SX;Wu ZY;Wang N
背景脊髓性肌萎缩症（SMA）是一种由运动神经元存活基因1（SMN1）突变引起的常染色体隐性遗传性疾病。最近，使用饱和 LC Green 染料进行高分辨率 DNA 熔解分析 (HRMA) 已成为用于基因分型或突变扫描的强大 PCR 后技术。目前尚未有研究将HRMA应用于SMA的分子分析。方法分别采用无标记探针的不对称PCR和无探针的对称PCR对55例SMA患者和46例无关正常人的SMN1和SMN2基因的外显子7和侧翼区进行扩增。将饱和 LC 绿色染料添加到 PCR 系统中。将 PCR 产物加载到 LightScanner 系统上，并从 60°C 缓慢熔化至 95°C。通过LightScanner软件获取并分析熔解曲线。结果在未标记探针的HRMA色谱图上，与SMA患者和对照的推测基因型相关的三种类型的熔解曲线清晰分离。对 55 名 SMA 患者和 46 名非 SMA 对照进行了 HRMA 鉴定，临床敏感性为 100%。 结论 采用饱和 LC Green 染料和未标记探针的 HRMA 似乎是诊断 SMA 的合适替代方法，具有高敏感性和特异性。
BackgroundSpinal muscular atrophy (SMA) is an autosomal recessive hereditary disorder caused by mutations of the survival motor neuron 1 (SMN1) gene. Recently, high-resolution DNA melting analysis (HRMA) with saturation LC Green dyes has become a powerful post-PCR technique for genotyping or mutation scanning. So far, no studies have applied HRMA to the molecular analysis of SMA.MethodsThe exon 7 and the flanking area of theSMN1andSMN2genes of 55 SMA patients and 46 unrelated normal individuals were amplified with asymmetric PCR with unlabeled probe and symmetric PCR without probe, respectively. The saturation LC Green dyes were added to the PCR system. The PCR products were loaded onto the LightScanner system and were melted from 60°C to 95°C slowly. The melting curves were acquired and analyzed by the LightScanner software.ResultsThree types of melting curves that correlated with the presumed genotype of SMA patients and controls were clearly separated on the HRMA chromatogram with the unlabeled probe. The 55 SMA patients and 46 non-SMA controls were identified with HRMA with a 100% clinical sensitivity.ConclusionThe HRMA with saturation LC Green dyes and unlabeled probe appears to be a suitable, alternative method for the diagnosis of SMA, with high sensitivity and specificity.
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影响因子：
64.5
作者：
LEFEBVRE, S;BURGLEN, L;MELKI, J
通讯作者：
MELKI, J
DOI：
10.1016/j.jenctecman.2005.09.003
发表时间：
2005-12-01
影响因子：
4.8
作者：
Chen, S
通讯作者：
Chen, S
影响因子：
1.9
作者：
Mazzei, R;Conforti, FL;Quattrone, A
通讯作者：
Quattrone, A
影响因子：
9.3
作者：
Pyatt, Robert E.;Mihal, David C.;Prior, Thomas W.
通讯作者：
Prior, Thomas W.
影响因子：
9.8
作者：
Feldkötter, M;Schwarzer, V;Wirth, B
通讯作者：
Wirth, B