A Burkholderia pseudomallei protein microarray reveals serodiagnostic and cross-reactive antigens
A Burkholderia pseudomallei protein microarray reveals serodiagnostic and cross-reactive antigens
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DOI：
10.1073/pnas.0812080106
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发表时间：
2009-08-11
影响因子：
11.1
通讯作者：
Titball, Richard W.
中科院分区：
文献类型：
--
作者：
Felgner, Philip L.;Kayala, Matthew A.;Titball, Richard W.
了解免疫系统对感染的反应方式对于疫苗和许多诊断方法的开发至关重要。为了深入了解这一领域，我们制作了一个含有1,205个类鼻疽伯克霍尔德菌蛋白的蛋白质微阵列，用88个类鼻疽患者血清对其进行探测，并鉴定了170个反应性抗原。将该抗原亚组打印在较小的阵列上，并用来自10个患者组的747份个体血清的集合进行探测，所述患者组包括来自泰国东北部和新加坡的类鼻疽患者、具有不同感染的患者、来自美国的健康个体以及来自泰国的地方性和非地方性地区的健康个体。我们确定了49种抗原，这些抗原在类鼻疽患者中的反应性明显高于健康人和其他类型细菌感染的患者。我们还鉴定了59种交叉反应性抗原，这些抗原在所有组中具有相同的反应性，包括来自美国的健康对照。利用这些结果，我们能够设计出一种测试，可以分类类鼻疽阳性和阴性个体，灵敏度和特异性分别为95%和83%，这是对目前可用的诊断测定的显著改进。一半的反应性抗原含有预测的信号肽序列，并被归类为外膜，表面结构或分泌分子，和额外的20%与致病性，适应或伴侣。这些结果表明，微阵列允许在抗原特异性、患者特异性和人群特异性的基础上对免疫应答进行更全面的分析，可以识别血清学诊断抗原，并有助于更详细地了解这种病原体的免疫原性。
Understanding the way in which the immune system responds to infection is central to the development of vaccines and many diagnostics. To provide insight into this area, we fabricated a protein microarray containing 1,205 Burkholderia pseudomallei proteins, probed it with 88 melioidosis patient sera, and identified 170 reactive antigens. This subset of antigens was printed on a smaller array and probed with a collection of 747 individual sera derived from 10 patient groups including melioidosis patients from Northeast Thailand and Singapore, patients with different infections, healthy individuals from the USA, and from endemic and nonendemic regions of Thailand. We identified 49 antigens that are significantly more reactive in melioidosis patients than healthy people and patients with other types of bacterial infections. We also identified 59 cross-reactive antigens that are equally reactive among all groups, including healthy controls from the USA. Using these results we were able to devise a test that can classify melioidosis positive and negative individuals with sensitivity and specificity of 95% and 83%, respectively, a significant improvement over currently available diagnostic assays. Half of the reactive antigens contained a predicted signal peptide sequence and were classified as outer membrane, surface structures or secreted molecules, and an additional 20% were associated with pathogenicity, adaptation or chaperones. These results show that microarrays allow a more comprehensive analysis of the immune response on an antigen-specific, patient-specific, and population-specific basis, can identify serodiagnostic antigens, and contribute to a more detailed understanding of immunogenicity to this pathogen.