羊布氏菌病iiELISA抗体检测方法的建立及应用

Establishment and Application of an Indirect ELISA for Brucella Antibody Detection in Goats and Sheep

关键词:  羊布鲁氏菌病, iELISA

Key words:  Goat  and sheep  Brucellosis, iELISA

摘要:

摘要：本研究建立了以布鲁氏菌LPS为抗原，用于检测羊布鲁氏菌IgG抗体的间接酶联免疫吸附试验（iELISA）方法。对该方法的敏感性、特异性、重复性、稳定性、符合率进行了评价。试验结果表明，iELISA比SAT和RBPT的敏感性高60-150倍；该方法特异性强，可有效区分布鲁氏菌与大肠杆菌O:157和都柏林沙门氏菌的IgG抗体，但尚无法与小肠结肠炎耶尔森菌O:9的IgG抗体相区分；批间重复性试验和批内重复性试验变异系数分别为1.1%-9.8%，2.9%-9.8%，表明该方法具有良好的可重复性；保存期试验显示包被好的酶标板在4℃保存11个月，检测结果稳定。iELISA和RBPT检测385份来自不同地区的血清结果表明，阳性符合率为95.14%，阴性符合率为96.28%，对1932份临床血清样本的检测结果与RBPT比较，总符合率为91.7%，证明此方法可以应用于临床羊布鲁氏菌病的筛查。

Abstract:

Abstract An indirect enzyme-linked immunosorbent assay based on Brucella LPS antigen was established for the detection of IgG antibodies against brucella in goats and sheep. The sensitivity, specificity, repeatability, stability and the coincidence of the iELISA were evaluated. Results showed that iELISA was 60-150 times more sensitive than Rose Bengal Plate Test and Standard Tube Agglutination Test. Specificity test showed that iELISA could effectively distinguish brucella LPS-IgG from that of Escherichia Coli O:157 and Salmonella Dublin, but could not distinguish from Yersinia enterocolitica LPS-IgG. The coefficient of variation of test results in batches and among batches were 1.1%-9.8%，2.9%-9.8% respectively, showing satisfied stability. The test results with the LPS-coated ELISA plates which were kept at 4 ℃ for 11 months, were stable. The coincidence test results of 385 serum from different areas showed that the positive coincidence was 95.14%, the negative coincidence was 96.28% and the whole coincidence of the 1932 clinical samples was 91.7%.. The evaluation results showed that iELISA established can be applied in the screen of IgG antibodies against brucella in goats and sheep.