Characterization of the main immunogenic proteins in Brucella infection for their application in diagnosis of brucellosis
Characterization of the main immunogenic proteins in Brucella infection for their application in diagnosis of brucellosis
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布鲁氏菌感染中主要免疫原性蛋白的表征及其在布鲁氏菌病诊断中的应用
DOI：
10.1016/j.cimid.2020.101462
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发表时间：
2020-06-01
影响因子：
2
通讯作者：
Yu, Shengqing
中科院分区：
文献类型：
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作者：
Tian, Mingxing;Song, Meiying;Yu, Shengqing
布鲁氏菌病是一种在世界范围内广泛传播的重要人畜共患细菌性疾病。控制该疾病的关键步骤是准确诊断和淘汰患病动物。经典的诊断方法，如试管凝集试验，由于与小肠结肠炎耶尔森菌O:9血清型存在交叉反应，不准确且不具特异性。此前，有几种蛋白质被报道为布鲁氏菌的主要免疫原。在本研究中，我们利用动物感染模型评估了针对外膜蛋白16（OMP16）、BP26、BLS、BCSP31、VirB12、超氧化物歧化酶C（SodC）和热休克蛋白60（GroEL）等蛋白质的抗体产生情况，并研究了它们在布鲁氏菌病诊断中的应用。结果表明，BP26和BLS是两种免疫原性最强的蛋白质。在进一步的研究中，我们用蛋白质印迹法检测了44份临床牛血清，结果显示BP26和BLS与30份布鲁氏菌阳性血清发生反应，但在14份无布鲁氏菌血清中也出现了假阳性结果。在间接酶联免疫吸附测定（ELISA）中，与基于脂多糖的ELISA相比，基于BP26的ELISA在布鲁氏菌阳性血清中的符合率为92.68%，但在无布鲁氏菌血清中仅为52.94%。基于BLS的ELISA几乎无法区分阳性血清和阴性血清。此外，BP26蛋白的截短片段在检测无布鲁氏菌血清时无法排除假阳性结果。总之，尽管布鲁氏菌的主要免疫原性蛋白质与布鲁氏菌阳性血清有良好的反应，但与无布鲁氏菌血清的假阳性反应可能导致布鲁氏菌病的误诊，这表明在将这些免疫原性蛋白质用作布鲁氏菌病诊断的抗原靶点时应更加谨慎。
Brucellosis is an important zoonotic bacterial disease widespread in the world. The key step of control this disease is accurate diagnosis and elimination of diseased animals. The classic diagnostic methods, such as tube agglutination test, are inaccurate and nonspecific, because of cross-reaction with Yersinia enterocolitica serotype O:9. Previously, several proteins were reported as Brucella main immunogens. In this study, we used animal infection model to evaluate antibody production against OMP16, BP26, BLS, BCSP31, VirB12, SodC and GroEL proteins and investigated their application in diagnosis of brucellosis. The results showed that the BP26 and BLS are two best immunogenic proteins. In further study, we detected 44 clinical bovine sera using western blot, showing that the BP26 and BLS reacted with 30 Brucella-positive sera, but false-positive results were also shown in 14 Brucella-free sera. In an indirect ELISA assay, compared to lipopolysaccharide-based ELISA, the conformance of the BP26-based ELISA was 92.68 % in Brucella-positive sera, but only 52.94 % in Brucella-free sera. The BLS-based ELISA can hardly differentiate positive sera from negative sera. Besides, truncated fragments of the BP26 protein cannot exclude false-positive results in detection of Brucella-free sera. Altogether, although Brucella main immunogenic proteins have good reaction with Brucella-positive sera, false-positive reaction with Brucella-free sera may lead to misdiagnosis of brucellosis, suggesting that it should be more careful to use these immunogenic proteins as antigen targets to diagnosis of brucellosis.