Assessment of liquid microbead arrays for the screening of newborns for spinal muscular atrophy
Assessment of liquid microbead arrays for the screening of newborns for spinal muscular atrophy
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DOI：
10.1373/clinchem.2007.092312
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发表时间：
2007-11-01
影响因子：
9.3
通讯作者：
Prior, Thomas W.
中科院分区：
文献类型：
--
作者：
Pyatt, Robert E.;Mihal, David C.;Prior, Thomas W.
背景资料：脊髓性肌萎缩症是一种常见的神经退行性疾病，最近已被考虑纳入下一代新生儿筛查方案。我们试图通过直接DNA分析来验证液体微珠阵列是否可用于识别受影响个体。方法：通过使用Luminex 200上的2种不同微珠化学物质，创建检测试剂盒来检测大约95%受影响个体中SMN 1基因外显子7的纯合缺失：MultiCode-PLx和Tag-It。然后使用每种检测方法分析一系列367个血液斑点，包括164个来自受影响个体，46个来自已知携带者，157个来自未受影响个体。MultiCode-PLx检测试剂盒需要4.2小时才能完成，并可正确鉴定来自受影响个体的所有164份样本。还对所有46份携带者和157份未受影响的个体样本进行了正确排除。Tag-It检测需要6.8小时，检测到所有受影响个体的样本，并排除了除1个（99.5%）样本外的所有样本。两种方法均不敏感增加拷贝数的SMN 2基因。结论：这两种方法显示出较高的敏感性和特异性检测脊髓性肌萎缩症患者。对于这两种方法，从所有血斑中提取大量DNA进行分析，SMN 2拷贝数不干扰。液体微珠阵列代表了新生儿筛查实验室中DNA分析的一种稳健方法。(C)2007年美国临床化学协会。
Background: Spinal muscular atrophy is a common neurodegenerative disorder that has recently been considered for inclusion in the next generation of newborn screening regimens. We sought to validate liquid microbead arrays for the identification of affected individuals by direct DNA analysis.Methods: Assays were created to detect the homozygous deletions in exon 7 of the SMN1 gene found in approximately 95% of affected individuals by use of 2 different microbead chemistries on the Luminex 200: MultiCode-PLx and Tag-It. A series of 367 blood spots including 164 from affected individuals, 46 from known carriers, and 157 from unaffected individuals were then analyzed with each assay.Results: The MultiCode-PLx assay required 4.2 h to perform and provided correct identification of all 164 samples from affected individuals. Correct exclusion was also made for all 46 carrier and 157 unaffected individual samples. The Tag-It assay required 6.8 h, detected all samples from affected individuals, and excluded all but 1 (99.5%) of the samples from carriers and unaffected individuals. Neither method was sensitive to increasing copy numbers of the SMN2 gene.Conclusions: Both methods showed high sensitivity and specificity for the detection of patients with spinal muscular atrophy. For both methods, ample DNA was extracted from all blood spots for analysis, and SMN2 copy numbers did not interfere. Liquid bead arrays represent a robust method for DNA analysis in newborn screening laboratories. (C) 2007 American Association for Clinical Chemistry.