Division of Infectious Diseases, Department of Medicine, University of California Irvine, Irvine, California, USA.
PLoS Negl Trop Dis. 2010 May 4;4(5):e673. doi: 10.1371/journal.pntd.0000673.
Brucellosis is a widespread zoonotic disease that is also a potential agent of bioterrorism. Current serological assays to diagnose human brucellosis in clinical settings are based on detection of agglutinating anti-LPS antibodies. To better understand the universe of antibody responses that develop after B. melitensis infection, a protein microarray was fabricated containing 1,406 predicted B. melitensis proteins. The array was probed with sera from experimentally infected goats and naturally infected humans from an endemic region in Peru. The assay identified 18 antigens differentially recognized by infected and non-infected goats, and 13 serodiagnostic antigens that differentiate human patients proven to have acute brucellosis from syndromically similar patients. There were 31 cross-reactive antigens in healthy goats and 20 cross-reactive antigens in healthy humans. Only two of the serodiagnostic antigens and eight of the cross-reactive antigens overlap between humans and goats. Based on these results, a nitrocellulose line blot containing the human serodiagnostic antigens was fabricated and applied in a simple assay that validated the accuracy of the protein microarray results in the diagnosis of humans. These data demonstrate that an experimentally infected natural reservoir host produces a fundamentally different immune response than a naturally infected accidental human host.
布鲁氏菌病是一种广泛流行的人畜共患病，也是生物恐怖主义的潜在制剂。目前，用于临床诊断人类布鲁氏菌病的血清学检测方法是基于检测抗 LPS 抗体的凝集反应。为了更好地了解感染 B. melitensis 后产生的抗体反应范围，我们制作了一个包含 1406 个预测的 B. melitensis 蛋白的蛋白质微阵列。用来自秘鲁流行地区的实验感染山羊和自然感染人类的血清探测该阵列。该检测方法鉴定出了 18 种被感染和未感染山羊差异识别的抗原，以及 13 种血清学诊断抗原，可区分患有急性布鲁氏菌病的人类患者和具有相似综合征的患者。在健康山羊中有 31 种交叉反应抗原，在健康人中则有 20 种。在人类和山羊中，只有两种血清学诊断抗原和八种交叉反应抗原重叠。基于这些结果，我们制作了包含人类血清学诊断抗原的硝酸纤维素线印迹，并应用于一种简单的检测方法，验证了蛋白质微阵列在人类诊断中的准确性。这些数据表明，实验感染的天然宿主产生的免疫反应与自然感染的偶然人类宿主产生的免疫反应根本不同。
