Yin Dehui, Li Li, Song Xiuling, Li Han, Wang Juan, Ju Wen, Qu Xiaofeng, Song Dandan, Liu Yushen, Meng Xiangjun, Cao Hongqian, Song Weiyi, Meng Rizeng, Liu Jinhua, Li Juan, Xu Kun
Department of Health Laboratory, School of Public Health, Jilin University, Changchun, China.
Department of Infection Control, First Hospital of Jilin University, Changchun, China.
BMC Infect Dis. 2016 May 21;16:219. doi: 10.1186/s12879-016-1552-9.
In epidemic regions of the world, brucellosis is a reemerging zoonosis with minimal mortality but is a serious public hygiene problem. Currently, there are various methods for brucellosis diagnosis, however few of them are available to be used to diagnose, especially for serious cross-reaction with other bacteria.
To overcome this disadvantage, we explored a novel multi-epitope recombinant protein as human brucellosis diagnostic antigen. We established an indirect enzyme-linked immunosorbent assay (ELISA) based on this recombinant protein. 248 sera obtained from three different groups including patients with brucellosis (146 samples), non-brucellosis patients (82 samples), and healthy individuals (20 samples) were tested by indirect ELISA. To evaluate the assay, a receiver-operating characteristic (ROC) analysis and immunoblotting were carried out using these characterized serum samples.
For this test, the area under the ROC curve was 0.9409 (95 % confidence interval, 0.9108 to 0.9709), and a sensitivity of 88.89 % and a specificity of 85.54 % was given with a cutoff value of 0.3865 from this ROC analysis. The Western blot results indicate that it is feasible to differentiate human brucellosis and non-brucellosis with the newly established method based on this recombinant protein.
Our results obtained high diagnostic accuracy of the ELISA assay which encourage the use of this novel recombinant protein as diagnostic antigen to implement serological diagnosis of brucellosis.
在世界流行地区,布鲁氏菌病是一种再度出现的人畜共患病,死亡率极低,但却是一个严重的公共卫生问题。目前,布鲁氏菌病有多种诊断方法,然而其中很少有可用于诊断的方法,尤其是对于与其他细菌的严重交叉反应。
为克服这一缺点,我们探索了一种新型多表位重组蛋白作为人类布鲁氏菌病诊断抗原。我们基于该重组蛋白建立了间接酶联免疫吸附测定(ELISA)法。用间接ELISA法检测了从三组不同人群中获得的248份血清,这三组人群包括布鲁氏菌病患者(146份样本)、非布鲁氏菌病患者(82份样本)和健康个体(20份样本)。为评估该检测方法,使用这些特征明确的血清样本进行了受试者工作特征(ROC)分析和免疫印迹分析。
对于该检测,ROC曲线下面积为0.9409(95%置信区间,0.9108至0.9709),根据该ROC分析,临界值为0.3865时,敏感性为88.89%,特异性为85.54%。蛋白质印迹结果表明,基于该重组蛋白的新建立方法区分人类布鲁氏菌病和非布鲁氏菌病是可行的。
我们的结果表明ELISA检测具有较高的诊断准确性,这鼓励使用这种新型重组蛋白作为诊断抗原,以实施布鲁氏菌病的血清学诊断。