2025年7月30日,Ivanka Krasteva等作为原著作者，在《Frontiers in Microbiology》发表“Identifying immunoreactive proteins in brucellin for enhanced brucellosis diagnosis: a proteomic approach”论文。
研究要点
布鲁菌素皮肤试验（BST）通过细胞介导的免疫反应检测动物布鲁菌病，该反应针对来自羊种布鲁菌115株的蛋白质提取物，该提取物几乎不含脂多糖。
该试验具有高度特异性，用于确认小反刍动物和猪中疑似假阳性血清学结果，但由于灵敏度低，不推荐用于筛查。
尽管布鲁菌素具有诊断意义，但其蛋白质组成尚未完全表征。
本研究使用nLC-ESIMS/MS分析和生物信息学工具评估布鲁菌素的蛋白质组成并鉴定免疫反应性蛋白质。
使用了由ANSES、IZS-Teramo（IZSAM）生产的欧盟标准布鲁菌素纯化蛋白质（不含S-LPS）的过敏原悬浮液以及先前商业化的布鲁菌素OCB®。
蛋白质组学分析鉴定出247种（ANSES）、542种（IZSAM）和183种（OCB）蛋白质。
其中206种蛋白质（ANSES）、458种蛋白质（IZSAM）和156种（OCB）被预测为潜在抗原，123种蛋白质在所有三种布鲁菌素中均存在。
在所有三种布鲁菌素制剂共有的123种蛋白质中，检测到先前在布鲁菌研究中已识别的几种关键免疫显性蛋白质——如核糖体L7/L12、外膜蛋白BP26/OMP28、GroEL和细菌铁蛋白。
它们在所有制剂中的存在支持了它们在诱导迟发型超敏反应和促进布鲁菌发病机制中的重要作用。
这些发现强调了引入质谱分析作为布鲁菌素批次生产质量控制的重要性，以及这些蛋白质作为检测布鲁菌细胞免疫候选物的潜力。
开发重组布鲁菌过敏原蛋白质有助于标准化皮肤试验，提供可靠的过敏原以促进疾病控制和根除。
此外，使用这些重组蛋白质的血清学检测可以提高当前布鲁菌间接试验的特异性，并消除与基于LPS的诊断相关的假阳性结果。
图文速览
Figure 1 Venn diagram illustrating the overlap of proteins identified by LC-MS/MS in three brucellins formulations: ANSES, IZSAM, and the commercial brucellergene OCB®. A total of 141 proteins were shared among all three formulations. In total, 247 proteins were identified in ANSES, 542 in IZSAM, and 183 in OCB
Figure 2 Overview of bioinformatics tools used for prediction of protein candidates
Figure 3 Venn diagram of three brucellin formulations (ANSES, IZSAM, and brucellergene OCB®), showing the overlap of immunogenic proteins identified by LC-MS/MS and predicted by VirulentPred and VaxiJen v2.0 (threshold >0.4). A total of 205 proteins in ANSES, 458 in IZSAM and 156 in OCB® were classified as potential antigens. Among these, 123 immunogenic proteins were common to all three formulations. Selected examples of these shared proteins, such as L7/L12, outer membrane protein BP26/OMP28, GroEL, and Bacterioferritin are indicated in the diagram. A complete list is provided in Supplementary Tables 1–3
Figure 4 Functional classification of the 123 proteins shared among Brucellin ANSES, Brucellin IZSM, and Brucellergene OCB® based on KEGG pathway annotation using BlastKOALA. The pie chart illustrates the distribution of proteins across functional categories and pathways, with the majority associated with environmental information processing, carbohydrate metabolism, and protein families involved in genetic and cellular processes
Table 1 List of non-homologs proteins to cross-reactive bacteria, (Pseudomonas aeruginosa,Bordetella bronchiseptica,Actinobacillus equuli, Streptococcus spp., Staphylococcus spp., Moraxella type, Salmonella spp., Campylobacter spp. and Yersinia enterocolitica O:9)
附件：
鉴定布鲁氏素中的免疫反应蛋白以增强布鲁氏菌病诊断：一项蛋白质组学方法.pdf
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