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Abstract：
Brucellosis is an economically devastating animal disease and has public health concern. Serological methods such as Rose Bengal Plate Test (RBPT), Complement Fixation Test (CFT), and Indirect-Enzyme-Linked Immunosorbent Assay (I-ELISA) have been used to detect brucellosis. However, there is limited comparative evaluation studies and lack of molecular confirmation of the causative agents in the study areas. The study was aimed to compare RBPT, I-ELISA, CFT, and confirmation using Polymerase Chain Reaction (PCR). A total of 2317 sera samples were collected from brucellosis-affected areas of Ethiopia with no vaccination history. All sera were subjected to comparative serological assays. Post-cross tabulation, sensitivity, and specificity were determined using Receiver Operating Characteristics (ROC) curve analysis software. PCR was performed on 54 seropositive samples using genus- and species-specific primers.
Among the 2317 sera tested for comparative serological assays, 189 (8.16%) were positive for RBPT, 191 (8.24%) for I-ELISA, and 48 (2.07%) for CFT. Sensitivity to RBPT was 100% (95%) in shoats and 74% (95%) in cattle. Specificity on RBPT was 98.69% (95%), 99.28% (95%), 100% (95%) in sheep, goats, and cattle, respectively. CFT sensitivity was 4 (95%) in sheep, 9.65 (95%) goats, and 72 (95%) cattle. Specificity on CFT was 100% (95%) for sheep, goats, and cattle. A 223bp Brucella genus-specific and 156bp B. abortus species-specific detected. However, B. melitensis not detected.
In this study, I-ELISA was the most sensitive and specific test. RBPT detected all Brucellosis-infected sheep and goats; nevertheless, it showed false positive in sheep and goats and false negative in cattle. The presence of B. abortus in small and large ruminants was confirmed by PCR. This is the first report of B. abortus detection in small ruminant in Ethiopia. B.abortus detected in non-preferred hosts. The findings suggest further study on molecular epidemiology of Brucella species.
Among the 2317 sera tested for comparative serological assays, 189 (8.16%) were positive for RBPT, 191 (8.24%) for I-ELISA, and 48 (2.07%) for CFT. Sensitivity to RBPT was 100% (95%) in shoats and 74% (95%) in cattle. Specificity on RBPT was 98.69% (95%), 99.28% (95%), 100% (95%) in sheep, goats, and cattle, respectively. CFT sensitivity was 4 (95%) in sheep, 9.65 (95%) goats, and 72 (95%) cattle. Specificity on CFT was 100% (95%) for sheep, goats, and cattle. A 223bp Brucella genus-specific and 156bp B. abortus species-specific detected. However, B. melitensis not detected.
In this study, I-ELISA was the most sensitive and specific test. RBPT detected all Brucellosis-infected sheep and goats; nevertheless, it showed false positive in sheep and goats and false negative in cattle. The presence of B. abortus in small and large ruminants was confirmed by PCR. This is the first report of B. abortus detection in small ruminant in Ethiopia. B.abortus detected in non-preferred hosts. The findings suggest further study on molecular epidemiology of Brucella species.
摘要：
背景：布鲁氏菌病是一种具有经济破坏性的动物疾病，具有公共卫生问题。血清学方法，如玫瑰红平板试验(RBPT)，补体固定试验(CFT),间接酶联免疫吸附测定(I-ELISA)已用于检测布鲁氏菌病。然而,在研究区域中，比较评估研究有限，缺乏对病原体的分子确认。这项研究的目的是比较RBPT，I-ELISA,CFT,并使用聚合酶链反应（PCR）进行确认。从埃塞俄比亚布鲁氏菌病感染地区收集了2317份血清样本，没有疫苗接种史。对所有血清进行比较血清学测定。交叉后制表，灵敏度,使用受试者工作特征(ROC)曲线分析软件确定和特异性。使用属和种特异性引物对54份血清阳性样品进行PCR。
结果：在用于比较血清学测定的2317血清中，189例（8.16%）RBPT阳性，191（8.24%）的I-ELISA，CFT为48（2.07%）。shoat对RBPT的敏感性为100％（95％），牛对RBPT的敏感性为74％（95％）。RBPT的特异性为98.69％（95％），99.28%（95%），100%(95%)的羊,山羊,和牛,分别。绵羊的CFT敏感性为4(95%)，9.65（95%）山羊，72头（95%）牛。绵羊CFT的特异性为100%(95%)，山羊,和牛。检测到223bp的布鲁氏菌属特异性和156bp的流产芽孢杆菌特异性。然而,B.melitensis未检测到。
结论：在这项研究中，I-ELISA是最敏感和特异的检测方法。RBPT检测到所有感染布鲁氏菌病的绵羊和山羊；然而，它在绵羊和山羊中显示出假阳性，在牛中显示出假阴性。通过PCR确认小反刍动物和大反刍动物中流产芽孢杆菌的存在。这是埃塞俄比亚小反刍动物流产B.abortus检测的第一份报告。在非首选宿主中检测到B.abortus。这些发现建议对布氏杆菌的分子流行病学进行进一步研究。
结果：在用于比较血清学测定的2317血清中，189例（8.16%）RBPT阳性，191（8.24%）的I-ELISA，CFT为48（2.07%）。shoat对RBPT的敏感性为100％（95％），牛对RBPT的敏感性为74％（95％）。RBPT的特异性为98.69％（95％），99.28%（95%），100%(95%)的羊,山羊,和牛,分别。绵羊的CFT敏感性为4(95%)，9.65（95%）山羊，72头（95%）牛。绵羊CFT的特异性为100%(95%)，山羊,和牛。检测到223bp的布鲁氏菌属特异性和156bp的流产芽孢杆菌特异性。然而,B.melitensis未检测到。
结论：在这项研究中，I-ELISA是最敏感和特异的检测方法。RBPT检测到所有感染布鲁氏菌病的绵羊和山羊；然而，它在绵羊和山羊中显示出假阳性，在牛中显示出假阴性。通过PCR确认小反刍动物和大反刍动物中流产芽孢杆菌的存在。这是埃塞俄比亚小反刍动物流产B.abortus检测的第一份报告。在非首选宿主中检测到B.abortus。这些发现建议对布氏杆菌的分子流行病学进行进一步研究。
