Mutations causing severe combined immunodeficiency: detection with a custom resequencing microarray
Mutations causing severe combined immunodeficiency: detection with a custom resequencing microarray
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DOI：
10.1097/gim.0b013e31818063bc
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发表时间：
2008-08-01
影响因子：
8.8
通讯作者：
Puck, Jennifer M.
中科院分区：
文献类型：
--
作者：
Lebet, Tonya;Chiles, Richard;Puck, Jennifer M.
目的：由于疾病基因的多样性和大量的致病突变，包括持续发现的独特突变，严重联合免疫缺陷的突变诊断是具有挑战性的。重测序微阵列可以促进突变检测，增加早期诊断婴儿的机会，以便通过造血干细胞移植进行最佳抢救。研究方法：在分析了累积的突变后，我们开发了一种定制的Affyssin基因芯片（R）微阵列，包括代表严重联合免疫缺陷病基因外显子和侧翼区域的探针。通过阵列与标准双脱氧基因组测序分析DNA样品。我们检测了X连锁IL 2 RG变异的男性及其母亲，以及IL 7 R、JAK 3和DCLRE 1C常染色体变异的患者和携带者。结果：新的，独特的严重联合免疫缺陷突变是常见的。95-98%的重测序阵列调用率超过了基因芯片产品的规格，并且检测到已知样品中的所有47个点突变，以及22个致病插入和缺失中的12个位点。每个基因都有特定的核苷酸，通常不能正确识别，必须从分析中排除;排除率范围从0.4%（半合子IL 2 RG）到9.2%（杂合子JAK 3）。结论：基因芯片重测序是一种很有前途的技术，可以检测已知和新的突变的严重联合免疫缺陷病突变诊断。探针序列和分析算法的未来定制可以增加准确识别核苷酸的数量。
Purpose: Mutation diagnosis of severe combined immunodeficiency is challenging because of the multiplicity of disease genes and large number of disease-causing mutations, including unique ones that continue to be found. A resequencing microarray could facilitate mutation detection, increasing the chance of diagnosing infants early for optimal rescue by hematopoietic stem cell transplantation. Methods: After analyzing cumulative mutations, We developed a custom Affymetrix GeneChip (R) microarray including probes representing exons and flanking regions of severe combined immunodeficiency disease genes. DNA samples were analyzed by array versus standard dideoxy genomic sequencing. We tested males and their mothers with X-linked IL2RG variants and patients and carriers with autosomal variants in IL7R, JAK3, and DCLRE1C. Results: New, unique severe combined immunodeficiency mutations are frequent. Resequencing array call rates of 95-98% exceeded GeneChip product specifications, and all of 47 point mutations in known samples were detected, as were the sites of 12 of 22 disease-causing insertions and deletions. Each gene had particular nucleotides that were often not called correctly and had to be excluded from analysis; exclusion rates ranged from 0.4% (hemizygous IL2RG) to 9.2% (heterozygous JAK3). Conclusion: Microarray resequencing is a promising technology for severe combined immunodeficiency mutation diagnosis that can detect both known and new mutations. Future customization of probe sequences and analysis algorithms Could increase the number of accurately called nucleotides.