Development of a routine newborn screening protocol for severe combined immunodeficiency
Development of a routine newborn screening protocol for severe combined immunodeficiency
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DOI：
10.1016/j.jaci.2009.04.007
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发表时间：
2009-09-01
影响因子：
14.2
通讯作者：
Routes, John M.
中科院分区：
文献类型：
--
作者：
Baker, Mei W.;Grossman, William J.;Routes, John M.
背景：严重联合免疫缺陷（SCID）的特征是功能性T细胞和B细胞的缺失。如果没有早期诊断和治疗，患有严重联合免疫缺陷的婴儿会在出生后一年内死于严重感染。目的：通过定量新生儿筛查（NBS）卡上干血斑（DBS）的T细胞受体切除环（TRECs）来确定检测新生儿严重联合免疫缺陷的可行性。方法：在去识别的NBS卡上从DBS中提取DNA，并使用实时定量PCR（RT-qPCR）来确定TREC的数量。阳性对照包括DBS从一个1周龄的T-B-NK+患者与SCID和全血标本选择性耗尽的幼稚T celles.Results：从5766去识别DBS的TRECs的平均值和中位数分别为827和708，每3.2毫米打孔（类似于3 μ L全血）。10份样本在初始分析时未能扩增TREC;除1份外，所有样本在复检时均显示正常TREC和β-肌动蛋白扩增。在SCID或幼稚T细胞耗尽的样品中均未检测到TRECs，尽管存在正常水平的β-actin.Conclusions：使用RT-qPCR从新生儿DBS提取的DNA中定量TRECs是一种高度敏感和特异的SCID筛选试验。该检测方法目前正在威斯康星州用于婴儿SCID的常规筛查。（J Allergy Clin Immunol 2009;124：522-7.）
Background: Severe combined immunodeficiency ( SCID) is characterized by the absence of functional T cells and B cells. Without early diagnosis and treatment, infants with SCID die from severe infections within the first year of life.Objective: To determined the feasibility of detecting SCID in newborns by quantitating T-cell receptor excision circles (TRECs) from dried blood spots (DBSs) on newborn screening (NBS) cards.Methods: DNA was extracted from DBSs on deidentified NBS cards, and real-time quantitative PCR (RT-qPCR) was used to determine the number of TRECs. Positive controls consisted of DBS from a 1-week-old T-B-NK+ patient with SCID and whole blood specimens selectively depleted of naive T cells.Results: The mean and median numbers of TRECs from 5766 deidentified DBSs were 827 and 708, respectively, per 3.2-mm punch (similar to 3 mu L whole blood). Ten samples failed to amplify TRECs on initial analysis; all but 1 demonstrated normal TRECs and beta-actin amplification on retesting. No TRECs were detected in either the SCID or naive T-cell-depleted samples, despite the presence of normal levels of beta-actin.Conclusions: The use of RT-qPCR to quantitate TRECs from DNA extracted from newborn DBSs is a highly sensitive and specific screening test for SCID. This assay is currently being used in Wisconsin for routine screening infants for SCID. (J Allergy Clin Immunol 2009;124:522-7.)