Eoh Hyungjin, Jeon Bo-Young, Kim Zhiyeol, Kim Seung-Cheol, Cho Sang-Nae
Department of Microbiology, College of Medicine, Yonsei University, 250 Seongsanno, Seodaemun-gu, Seoul 120-752, Republic of Korea.
J Vet Diagn Invest. 2010 Jul;22(4):524-30. doi: 10.1177/104063871002200405.
Brucella abortus is a bacterium of brucellosis causing abortion in cattle. The diagnosis of bovine brucellosis mainly relies on serologic tests using smooth lipopolysaccharide (S-LPS) from B. abortus. However, the usefulness of this method is limited by false-positive reactions due to cross-reaction with other Gram-negative bacteria. In the present study, the eryC gene encoding B. abortus d-erythrulose 1-phosphate dehydrogenase, which is involved in the erythritol metabolism in virulent B. abortus strain but is absent from a B. abortus vaccine strain (S19), was cloned. Recombinant EryC was expressed and purified for the evaluation as a diagnostic reagent for bovine brucellosis. Other B. abortus proteins, Omp16, PP26, and CP39 were also purified and their seroreactivities were compared. Recombinant EryC, Omp16, PP26, and PP39 were all reactive to B. abortus-positive serum. The specificity of recombinant Omp16, PP26, CP39, and EryC, were shown to be approximately 98%, whereas that of B. abortus whole cell lysates was shown to be 95%. The sensitivity of Omp16, PP26, CP39, and EryC were 10%, 51%, 64%, and 43%, respectively, whereas that of B. abortus whole cell lysates was 53%. These results suggested that B. abortus EryC would be a potential reagent for diagnosis for bovine brucellosis as a single protein antigen.
牛流产布鲁氏菌是一种可导致牛流产的布鲁氏菌病细菌。牛布鲁氏菌病的诊断主要依靠使用来自牛流产布鲁氏菌的光滑脂多糖（S-LPS）进行血清学检测。然而，由于与其他革兰氏阴性菌发生交叉反应，这种方法的有效性受到假阳性反应的限制。在本研究中，编码牛流产布鲁氏菌d-赤藓酮糖1-磷酸脱氢酶的eryC基因被克隆，该酶参与强毒株牛流产布鲁氏菌的赤藓醇代谢，但在牛流产布鲁氏菌疫苗株（S19）中不存在。重组EryC被表达并纯化，用于评估其作为牛布鲁氏菌病诊断试剂的性能。其他牛流产布鲁氏菌蛋白，Omp16、PP26和CP39也被纯化，并比较了它们的血清反应性。重组EryC、Omp16、PP26和PP39均与牛流产布鲁氏菌阳性血清发生反应。重组Omp16、PP26、CP39和EryC的特异性约为98%，而牛流产布鲁氏菌全细胞裂解物的特异性为95%。Omp16、PP26、CP39和EryC的敏感性分别为10%、51%、64%和43%，而牛流产布鲁氏菌全细胞裂解物的敏感性为53%。这些结果表明，牛流产布鲁氏菌EryC作为单一蛋白抗原，有望成为牛布鲁氏菌病诊断的试剂。
