Faculty of Medical and Health Sciences, University of Auckland, Auckland, New Zealand; Maurice Wilkins Centre for Biodiscovery, University of Auckland, Auckland, New Zealand.
Faculty of Medical and Health Sciences, University of Auckland, Auckland, New Zealand.
J Infect. 2018 Apr;76(4):361-368. doi: 10.1016/j.jinf.2017.12.008. Epub 2017 Dec 18.
Streptococcal serology provides evidence of prior Group A Streptococcus (GAS) exposure, crucial to the diagnosis of acute rheumatic fever (ARF) and post-streptococcal glomerulonephritis. However, current tests, which measure anti-streptolysin-O and anti-DNaseB antibodies, are limited by false positives in GAS endemic settings, and incompatible methodology requiring the two tests to be run in parallel. The objective was to improve streptococcal serology by combining the novel GAS antigen, SpnA, with streptolysin-O and DNaseB in a contemporary, bead-based immunoassay.
Recombinant streptolysin-O, DNAseB and SpnA were conjugated to polystyrene beads with unique fluorescence positions so antibody binding to all three antigens could be detected simultaneously by cytometric bead array. Multiplex assays were run on sera collected in three groups: ARF; ethnically matched healthy children; and healthy adults.
The ability of the antigens to detect a previous GAS exposure in ARF was assessed using the 80th centile of the healthy children group as cut-off (upper limit of normal). SpnA had the highest sensitivity at 88%, compared with 75% for streptolysin-O and 56% for DNaseB.
SpnA has favorable immunokinetics for streptococcal serology, and can be combined with anti-streptolysin-O and anti-DNaseB in a multiplex format to improve efficiency and accuracy.
链球菌血清学提供了 A 组链球菌(GAS)既往暴露的证据,这对急性风湿热(ARF)和链球菌后肾小球肾炎的诊断至关重要。然而,目前的检测方法,即测量抗链球菌溶血素-O 和抗 DNA 酶 B 抗体,在 GAS 流行地区存在假阳性的局限性,并且由于方法学不兼容,需要同时进行这两种检测。本研究旨在通过将新型 GAS 抗原 SpnA 与链球菌溶血素-O 和 DNA 酶 B 结合到当代的基于微球的免疫分析中,来改进链球菌血清学。
将重组链球菌溶血素-O、DNA 酶 B 和 SpnA 与聚苯乙烯微球偶联,具有独特的荧光位置,因此通过流式细胞术微球分析可以同时检测到针对所有三种抗原的抗体结合。在三组血清中进行了多重检测:ARF 组;与之匹配的种族健康儿童组;和健康成人组。
使用健康儿童组第 80 百分位数作为截断值(正常值上限)来评估抗原在 ARF 中检测既往 GAS 暴露的能力。SpnA 的敏感性最高,为 88%,而链球菌溶血素-O 为 75%,DNA 酶 B 为 56%。
SpnA 具有有利的链球菌血清学免疫动力学特性,并且可以与抗链球菌溶血素-O 和抗 DNA 酶 B 结合在多重格式中,以提高效率和准确性。