Detection of immunoglobulin isotypes from dried blood spots.
Detection of immunoglobulin isotypes from dried blood spots.
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DOI：
10.1016/j.jim.2013.12.001
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发表时间：
2014-02
影响因子：
2.2
通讯作者：
Lawrence DA
中科院分区：
文献类型：
--
作者：
Andersen NJ;Mondal TK;Preissler MT;Freed BM;Stockinger S;Bell E;Druschel C;Louis GM;Lawrence DA
该研究旨在确定从干血斑（DBS）中恢复免疫球蛋白（Ig）同种型（IgG亚类，IgA， IgE和IgM类）的敏感性和可重复性，这是Upstate KIDS研究的一个方法组成部分。IgG1/2/3/4、IgA和IgM分析采用多路Luminex法；ELISA法检测IgE。去识别患者的血浆样本用于比较Luminex测定法和浊度法，浊度法通常用于定量临床样本中的IgA、IgG和IgM。将IgE ELISA法与免疫荧光法进行比较。在评估新生儿干血斑（NDBSs）的冲孔之前，比较了脐带血DBSs （CBDBSs）冲孔与来自相同脐带血的血浆的Ig回收率。虽然血浆和DBS中Ig的回收率不具有可比性，这可能是由于DBS样品中的细胞裂解物造成的，但分析是可重复的。此外，从CBDBSs中恢复的IgA、IgG2、IgG4和IgM水平与血浆中的水平呈正相关。脑起搏器数据是一个相对值，因为它不等同于血浆浓度。Upstate KIDs研究中108名新生儿的大部分Ig浓度都在新生儿血浆Ig水平范围内，除了IgG3。IgG4值变化幅度最大（0.01 ~ 319 mg/dl）， IgG1值变化幅度最小（85.2 ~ 960.4 mg/dl）。
The study was designed to determine the sensitivity and reproducibility of recovering immunoglobulin (Ig) isotypes (IgG subclasses, IgA, IgE and IgM classes) from dried blood spots (DBS), a methodologic subcomponent of the Upstate KIDS Study. A multiplexed Luminex assay was used for IgG1/2/3/4, IgA and IgM analysis; an ELISA was used for IgE. Plasma samples from de-identified patients were used to compare the Luminex assay with nephelometry, which is routinely used to quantify IgA, IgG and IgM in clinical samples. The IgE ELISA was compared to an immunofluorescence assay. Prior to evaluation of punches from newborn dried blood spots (NDBSs), recoveries of Ig from punches of cord blood DBSs (CBDBSs) vs. plasma from the same cord bloods were compared. Although the recoveries of Ig from plasma and DBSs were not comparable, which could be due to cell lysates in the DBS samples, the analyses were reproducible. Additionally, the levels of IgA, IgG2, IgG4, and IgM recovered from CBDBSs positively correlated with those in plasma. The DBS data is a relative value since it is not equivalent to the plasma concentration. The majority of Ig concentrations recovered from 108 newborns of the Upstate KIDs Study were within the range of newborn plasma Ig levels with the exception of IgG3. The IgG4 values displayed the greatest variance with a wide range (0.01–319 mg/dl), whereas, IgG1 values had the narrowest range (85.2–960.4 mg/dl).