Microarray screening of Guillain-Barré syndrome sera for antibodies to glycolipid complexes.
Microarray screening of Guillain-Barré syndrome sera for antibodies to glycolipid complexes.
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DOI：
10.1212/nxi.0000000000000284
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发表时间：
2016-12
期刊：
影响因子：
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通讯作者：
Willison HJ
中科院分区：
文献类型：
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作者：
Halstead SK;Kalna G;Islam MB;Jahan I;Mohammad QD;Jacobs BC;Endtz HP;Islam Z;Willison HJ
利用一种新开发的微阵列技术，研究在孟加拉国收集的大量格林-巴利综合征(GBS)和对照样本中糖脂复合体自身抗体的模式。使用微阵列打印机将12种通常研究过的糖脂和脂类，以及它们的66种可能的异构体复合体，总计78个抗原，应用于聚偏氟乙烯涂布的玻片上。用266GBS和579份对照血清(每个血清2μL，稀释1/50)和结合免疫球蛋白G用二抗检测。对扫描的阵列进行统计分析。检测单靶抗体的敏感性比检测异构体复合靶抗体低9%(49.2%比58.3%)，而对特异性(83.9%-85.0%)无显著影响。GBS血清的最佳筛选方法包括10种糖脂(4种单一糖脂GM1、GA1、GD1a、GQ1b及其6种异构体复合体)，总体检测灵敏度为64.3%，特异性为77.1%。值得注意的异构体靶标是GM1：GD1a、GM1：GQ1b和GA1：GD1a，其中观察到与络合物的排他性结合。通过将筛选平台微型化为微阵列平台，并应用简单的生物信息学来确定靶标的最佳灵敏度和特异性，可以实现合理的筛选方案以捕获糖脂复合体的巨大多样性。糖脂复合体是自身免疫性神经病病例中糖脂抗原的一个重要类别，需要特定的分析和生物信息学方法才能进行最佳检测。
To characterize the patterns of autoantibodies to glycolipid complexes in a large cohort of Guillain-Barré syndrome (GBS) and control samples collected in Bangladesh using a newly developed microarray technique. Twelve commonly studied glycolipids and lipids, plus their 66 possible heteromeric complexes, totaling 78 antigens, were applied to polyvinylidene fluoride–coated slides using a microarray printer. Arrays were probed with 266 GBS and 579 control sera (2 μL per serum, diluted 1/50) and bound immunoglobulin G detected with secondary antibody. Scanned arrays were subjected to statistical analyses. Measuring antibodies to single targets was 9% less sensitive than to heteromeric complex targets (49.2% vs 58.3%) without significantly affecting specificity (83.9%–85.0%). The optimal screening protocol for GBS sera comprised a panel of 10 glycolipids (4 single glycolipids GM1, GA1, GD1a, GQ1b, and their 6 heteromeric complexes), resulting in an overall assay sensitivity of 64.3% and specificity of 77.1%. Notable heteromeric targets were GM1:GD1a, GM1:GQ1b, and GA1:GD1a, in which exclusive binding to the complex was observed. Rationalizing the screening protocol to capture the enormous diversity of glycolipid complexes can be achieved by miniaturizing the screening platform to a microarray platform, and applying simple bioinformatics to determine optimal sensitivity and specificity of the targets. Glycolipid complexes are an important category of glycolipid antigens in autoimmune neuropathy cases that require specific analytical and bioinformatics methods for optimal detection.