Laimis Silimavicius, Lieve Tchebotarev, Mindaugas Zaveckas, Raimundas Razanskas, Laima Cepulyte, Karolina Bielske, Indre Kucinskaite-Kodze, Linas Griguola, Kotryna Linauskiene, Rasa Petraityte-Burneikiene
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引用次数: 0
Abstract
Background: Timothy grass (Phleum pratense) is a significant source of allergens, and recombinant allergens are increasingly used for diagnostic purposes. However, the performance of different recombinant allergen production systems in diagnostic assays needs further investigation to optimize their use in clinical settings.
Objective: The main objective of this study was to analyze and compare the diagnostic performance of recombinant timothy grass allergens produced in E. coli and N. benthamiana using a custom-made microarray chip.
Methods: Recombinant timothy grass allergens Phl p 1, Phl p 2, Phl p 5, Phl p 6, Phl p 11, and Phl p 12 were produced in E. coli and/or N. benthamiana. A total of 113 patient serum samples were tested to evaluate the diagnostic sensitivity, specificity, inter-assay variability, and correlation of allergen-specific IgE detection compared to commercial multiplex tests (ALEX and ISAC). Additionally, the prevalence of sIgE to these allergens was assessed.
Results: Phl p 1, Phl p 2, Phl p 5, Phl p 6 and Phl p 11 showed high or very high positive correlation in immunoreactivity with other commercial multiplex tests. Notably, Phl p 11 fused with maltose-binding protein (MBP) demonstrated high diagnostic specificity and sensitivity, with a 0.3 arbitrary cut-off value. However, a high intra-assay variation was observed. The study also assessed specific IgE prevalence to timothy grass allergens within the tested patient cohort.
Conclusions: Recombinant allergens from both E. coli and N. benthamiana demonstrated strong diagnostic potential on the microarray platform, with Phl p 11 (MBP-fused) showing particularly high performance. High intra-assay variation highlights the need for further optimization in allergen formulation and microarray storage conditions. These results highlight the potential of recombinant allergens for diagnostic applications, despite challenges with allergen stability in microarray formats. Specific IgE prevalence to timothy allergens revealed a sensitization profile consistent with findings from multiple studies.
背景：提摩西草（Phleum pratense）是一种重要的过敏原来源，重组过敏原越来越多地被用于诊断目的。然而，不同重组过敏原生产系统在诊断测定中的表现需要进一步研究，以优化其在临床环境中的应用：本研究的主要目的是使用定制的微阵列芯片分析和比较在大肠杆菌和N.benthamiana中生产的重组梯牧草过敏原的诊断性能：方法：重组梯牧草过敏原 Phl p 1、Phl p 2、Phl p 5、Phl p 6、Phl p 11 和 Phl p 12 是在大肠杆菌和/或 N. benthamiana 中产生的。共检测了 113 份患者血清样本，以评估过敏原特异性 IgE 检测与商业多重检测（ALEX 和 ISAC）相比的诊断灵敏度、特异性、检测间变异性和相关性。此外，还评估了这些过敏原特异性 IgE 的流行率：结果：Phl p 1、Phl p 2、Phl p 5、Phl p 6 和 Phl p 11 与其他商用多重检测方法的免疫反应呈高度或极高度正相关。值得注意的是，与麦芽糖结合蛋白（MBP）融合的 Phl p 11 显示出较高的诊断特异性和灵敏度，任意临界值为 0.3。然而，在检测中也发现了很大的差异。该研究还评估了接受测试的患者群中对梯牧草过敏原的特异性 IgE 患病率：结论：来自大肠杆菌和N. benthamiana的重组过敏原在微阵列平台上表现出很强的诊断潜力，其中Phl p 11（MBP融合）表现尤为突出。测定内的高度差异突出表明需要进一步优化过敏原配方和微阵列储存条件。这些结果凸显了重组过敏原在诊断应用中的潜力，尽管在微阵列格式中过敏原的稳定性面临挑战。貓尾草過敏原的特異性 IgE 發現與多項研究結果一致。
期刊介绍：
BMC Biotechnology is an open access, peer-reviewed journal that considers articles on the manipulation of biological macromolecules or organisms for use in experimental procedures, cellular and tissue engineering or in the pharmaceutical, agricultural biotechnology and allied industries.
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