Next-generation sequencing for mitochondrial diseases: A wide diagnostic spectrum
Next-generation sequencing for mitochondrial diseases: A wide diagnostic spectrum
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DOI：
10.1111/j.1442-200x.2012.03644.x
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发表时间：
2012-10-01
影响因子：
1.4
通讯作者：
Hahn, Si Houn
中科院分区：
文献类型：
--
作者：
Vasta, Valeria;Merritt, J. Lawrence, II;Hahn, Si Houn
背景资料：目前线粒体疾病的诊断方法需要侵入性程序，如肌肉活检和多种生化检测，但结果往往是不确定的。临床测序测试仅适用于有限数量的基因。最近，大规模平行测序已成为一个强大的工具，用于测试遗传异质性条件，如线粒体疾病。研究方法：对26名已知或疑似线粒体疾病的患者进行了靶向下一代测序，使用溶液中捕获908个已知和候选核基因的外显子和Illumina基因组分析仪。结果如下：18例各种异常呼吸链复合体（RCC）活动的患者中，除了SURF 1已知突变的参考对照样本外，线粒体RCC酶的亚基或组装因子均无分子缺陷。相反，在已知的致病基因包括CPT 2，POLG，PDSS 1，UBE 3A，SDHD，和一些潜在的致病性变异的候选基因，如MTO 1或SCL 7A 13的几个变体被确定。结论：仅测序RCC亚基和组装因子的核基因可能无法为疑似线粒体疾病患者提供诊断答案。目前的研究结果表明，线粒体疾病的诊断谱比以前认为的要广泛得多，这可能会导致误诊和/或不适当的治疗。总体分析灵敏度和精密度对于临床检测而言似乎是可接受的。尽管在所有患者中发现突变存在局限性，但本研究结果强调了靶向下一代测序的显著临床益处，并作为在该临床异质性患者组中扩展临床评价的原型。
Background: The current diagnostic approach for mitochondrial disorders requires invasive procedures such as muscle biopsy and multiple biochemical testing but the results are often inconclusive. Clinical sequencing tests are available only for a limited number of genes. Recently, massively parallel sequencing has become a powerful tool for testing genetically heterogeneous conditions such as mitochondrial disorders. Methods: Targeted next-generation sequencing was performed on 26 patients with known or suspected mitochondrial disorders using in-solution capture for the exons of 908 known and candidate nuclear genes and an Illumina genome analyzer. Results: None of the 18 patients with various abnormal respiratory chain complex (RCC) activities had molecular defects in either subunits or assembly factors of mitochondrial RCC enzymes except a reference control sample with known mutations in SURF1. Instead, several variants in known pathogenic genes including CPT2, POLG, PDSS1, UBE3A, SDHD, and a few potentially pathogenic variants in candidate genes such as MTO1 or SCL7A13 were identified. Conclusions: Sequencing only nuclear genes for RCC subunits and assembly factors may not provide the diagnostic answers for suspected patients with mitochondrial disorders. The present findings indicate that the diagnostic spectrum of mitochondrial disorders is much broader than previously thought, which could potentially lead to misdiagnosis and/or inappropriate treatment. Overall analytic sensitivity and precision appear acceptable for clinical testing. Despite the limitations in finding mutations in all patients, the present findings underscore the considerable clinical benefits of targeted next-generation sequencing and serve as a prototype for extending the clinical evaluation in this clinically heterogeneous patient group.