A multiplex immunoassay using the Guthrie specimen to detect T-cell deficiencies including severe combined immunodeficiency disease.
A multiplex immunoassay using the Guthrie specimen to detect T-cell deficiencies including severe combined immunodeficiency disease.
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DOI：
10.1373/clinchem.2010.144329
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发表时间：
2010-09
影响因子：
9.3
通讯作者：
Pass KA
中科院分区：
文献类型：
--
作者：
Janik DK;Lindau-Shepard B;Comeau AM;Pass KA
严重联合免疫缺陷(SCID)符合新生儿筛查小组的许多要求。两项新生儿筛查SCID的初步研究目前正在进行中，使用的是T细胞受体切除环(TREC)分析，这是一种分子技术。在这里，我们描述了一种免疫分析方法，用CD3作为T细胞的标志，CD45作为总白细胞的标志，可以用来检测格思里标本。使用了Luminex平台的多路复用功能。抗体对被用来捕获和检测从单个3 mm冲压的格思里标本中的CD3和CD45。每个生物标记的分析在相同的缓冲液中单独进行，然后结合起来创建多重分析。用已知数量的白细胞制成的校准剂，对CD_3和CD_(45)的检出限分别为0.25×10~6个/毫升和0.125×10~6个/毫升。亲和力试验表明，CD3和CD45抗体之间没有交叉反应。多重分析针对8个已知临床状态的编码样本进行了验证，并与鉴定它们的TREC检测结果相关联。CD345鉴定结果均正确。CD345检测的性能参数符合免疫分析普遍接受的性能特征。我们对阳性标本的检测分类与以前的TREC结果一致。CD345检测可作为TREC检测的一个可行的替代或补充，作为一种主要的筛查工具，用于检测格思里标本中的T细胞免疫缺陷，包括SCID。
Severe combined immunodeficiency (SCID) fulfills many of the requirements for addition to a newborn screening panel. Two newborn screening SCID pilot studies are now underway using the T-cell receptor excision circle (TREC) assay, a molecular technique. Here we describe an immunoassay with CD3 as a marker for T cells and CD45 as a marker for total leukocytes that can be used with the Guthrie specimen. The multiplexing capabilities of the Luminex platform were used. Antibody pairs were used to capture and detect CD3 and CD45 from a single 3-mm punch of the Guthrie specimen. The assay for each bio-marker was developed separately in identical buffers and then combined to create a multiplex assay. Using calibrators made from known amounts of leukocytes, a detection limit of 0.25 × 106 cells/mL for CD3 and 0.125 × 106 cells/mL for CD45 was obtained. Affinity tests showed no cross-reactivity between the antibodies to CD3 and CD45. The multiplex assay was validated against 8 coded specimens of known clinical status and linked to results from the TREC assay that had identified them. All were correctly identified by the CD345 assay. The performance parameters of the CD345 assay met the performance characteristics generally accepted for immunoassays. Our assay classifications of positive specimens concur with previous TREC results. This CD345 assay warrants evaluation as a viable alternative or complement to the TREC assay as a primary screening tool for detecting T-cell immunodeficiencies, including SCID, in Guthrie specimens.
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影响因子：
14.2
作者：
Chan, K;Puck, JM
通讯作者：
Puck, JM
影响因子：
158.5
作者：
Rieux-Laucat, F;Hivroz, C;Le Deist, F
通讯作者：
Le Deist, F
影响因子：
120.7
作者：
Routes, John M.;Grossman, William J.;Baker, Mei W.
通讯作者：
Baker, Mei W.
影响因子：
9.3
作者：
Gerstel-Thompson, Jacalyn L.;Wilkey, Jonathan F.;Comeau, Anne Marie
通讯作者：
Comeau, Anne Marie
影响因子：
9.3
作者：
Lindau-Shepard, Barbara A.;Pass, Kenneth A.
通讯作者：
Pass, Kenneth A.