A rapid, noninvasive immunoassay for frataxin: utility in assessment of Friedreich ataxia.
A rapid, noninvasive immunoassay for frataxin: utility in assessment of Friedreich ataxia.
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DOI：
10.1016/j.ymgme.2010.07.001
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发表时间：
2010-10
影响因子：
3.8
通讯作者：
Lynch, David R.
中科院分区：
文献类型：
--
作者：
Deutsch, Eric C.;Santani, Avni B.;Perlman, Susan L.;Farmer, Jennifer M.;Stolle, Catherine A.;Marusich, Michael F.;Lynch, David R.
弗里德赖希共济失调（FRDA）是一种常染色体隐性神经退行性疾病，由线粒体蛋白frataxin的减少引起。研究中的Frataxin水平通常通过患者成纤维细胞、淋巴细胞或肌肉活检的Western blot分析来测量；这些都不是在大规模临床研究中快速检测的理想方法。最近，一种快速、无创的侧流免疫分析法被开发出来，以准确测量frataxin蛋白的picogram水平，并被证明可以区分FRDA携带者、患者和对照组的淋巴母细胞样细胞。我们将免疫分析扩展到直接测量颊细胞和全血中的卵黄蛋白，这些细胞和全血来自大量的对照组、已知的携带者和临床试验人群的典型患者。在颊细胞中的试验与先前在淋巴母细胞样细胞中进行的研究具有相似程度的变异性（对照组的变异系数约为10%）。对照组、携带者和患者颊细胞的平均组值（分别为对照组的100、50.2和20.9%）和全血中提取的蛋白（分别为100、75.3和32.2%）之间的frataxin蛋白含量存在显著差异，尽管两组之间存在一些重叠。此外，frataxin水平与GAA重复序列长度呈负相关，与发病年龄直接相关。具有扩增GAA重复序列和鉴定出的frataxin点突变的受试者也在疾病范围内携带frataxin水平。一些显示FRDA表型但只携带单一可识别突变的患者FRDA患者范围内的frataxin水平。根据FXN基因的多重连接依赖探针扩增（multiplex lig- dependent probe amplification， MLPA）分析，该组的一名患者有一个新的缺失，包括FXN基因的外显子2和3。在临床试验等特殊情况下，侧流免疫分析可能是一种有用的非侵入性评估FRDA患者重复frataxin水平的有效方法，并且可以作为辅助诊断工具，帮助识别和表征非典型患者。
Friedreich ataxia (FRDA) is an autosomal recessive neurodegenerative disorder caused by reduced amounts of the mitochondrial protein frataxin. Frataxin levels in research studies are typically measured via Western blot analysis from patient fibroblasts, lymphocytes, or muscle biopsies; none of these is ideal for rapid detection in large scale clinical studies. Recently, a rapid, non-invasive lateral-flow immunoassay was developed to accurately measure picogram levels of frataxin protein and shown to distinguish lymphoblastoid cells from FRDA carriers, patients and controls. We expanded the immunoassay to measure frataxin directly in buccal cells and whole blood from a large cohort of controls, known carriers and patients typical of a clinical trial population. The assay in buccal cells shared a similar degree of variability with previous studies conducted in lymphoblastoid cells (~10% coefficient of variation in controls). Significant differences in frataxin protein quantity were seen between the mean group values of controls, carriers, and patient buccal cells (100, 50.2, and 20.9% of control, respectively) and in protein extracted from whole blood (100, 75.3, and 32.2%, respectively), although there was some overlap between the groups. In addition, frataxin levels were inversely related to GAA repeat length and correlated directly with age of onset. Subjects with one expanded GAA repeat and an identified frataxin point mutation also carried frataxin levels in the disease range. Some patients displaying an FRDA phenotype but carrying only a single identifiable mutation had frataxin levels in the FRDA patient range. One patient from this group has a novel deletion that included exons 2 and 3 of the FXN gene based on multiplex ligation-dependent probe amplification (MLPA) analysis of the FXN gene. The lateral flow immunoassay may be a useful means to non-invasively assess frataxin levels repetitively with minimal discomfort in FRDA patients in specific situations such as clinical trials, and as a complementary diagnostic tool to aid in identification and characterization of atypical patients.
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