■布鲁氏菌病的诊断很大程度上依赖于虎红平板凝集试验(RBPT)。然而,很难区分自然感染抗体阳性和疫苗接种抗体阳性,然而,鉴定特定的布鲁氏菌属自然感染。
■这里,我们分析了主要外膜蛋白(OMPs)的结构,OMP25和OMP31来自布鲁氏菌(B.Ovis)和melitensis布鲁氏菌(B.melitensis),这是绵羊布鲁氏菌病的主要病原体,发现OMP25和OMP31可以用作B.ovis和B.melitensis抗体的差异抗原。然后,我们表达了来自B.ovis的OMP25(OMP25o)和来自B.melitensis的OMP31(OMP31m)。
■它们在接种疫苗的绵羊血清的抗体检测中具有同等的效率,与RBPT结果一致。然而,通过流行病学调查,我们发现一些RBPT阳性样本通过基于OMP31m的血清抗体检测为阴性,但是这些样品通过OMP25o给出了阳性结果。我们通过基于B.ovis和B.melitensis特异性引物的PCR检测验证了这些OMP31m阴性但OMP25o阳性的样品,所有这些样本都是B.melitensis阴性。然而,6个样本中有4个是双歧杆菌阳性.这些结果表明,我们可以使用OMP25o和OMP31m抗体来诊断绵羊布鲁氏菌病,尤其是鉴别念珠菌的感染.
■目前,中国尚未批准基于B.ovis的疫苗,而B.ovis阳性样品应自然感染。吉林省应该有一定的隐性传播。应进行进一步的流行病学调查以监测自然感染。
The diagnosis of brucellosis largely relies on tiger red plate agglutination test (RBPT). However, it is difficult to distinguish between natural infection antibody positive and vaccination antibody positive, nevertheless, the identification of specific Brucella species natural infection.
Here, we analyzed the structure of main outer membrane proteins (OMPs), OMP25 and OMP31 from Brucella ovis (B. ovis) and Brucella melitensis (B. melitensis), which are the main pathogens of sheep brucellosis, and found the OMP25 and OMP31 could be used as the differential antigens for B. ovis and B. melitensis antibody. Then we expressed the OMP25 from B. ovis (OMP25o) and OMP31 from B. melitensis (OMP31m).
They have equally efficiency in antibody detection of vaccinated sheep serum, consistent with the RBPT results. However, through epidemiological investigations, we found some RBPT positive samples were negative by the OMP31m based serum antibody detection, but these samples gave positive results by the OMP25o. We verified these OMP31m negative but OMP25o positive samples by B. ovis and B. melitensis specific primers based PCR detection, and all these samples were B. melitensis negative. However, four out of six samples are B. ovis positive. These results showed that we could use the OMP25o and OMP31m to diagnose sheep brucellosis antibody, especially to discriminate the infection of the B. ovis.
Currently, China has not yet approved a vaccine based on B. ovis and B. ovis positive samples should be naturally infected. There should be some implicit transmission of B. ovis in Jilin province. Further epidemiological investigation should be conducted to monitor the B. ovis natural infection.