Díaz-Aparicio E, Marín C, Alonso-Urmeneta B, Aragón V, Pérez-Ortiz S, Pardo M, Blasco J M, Díaz R, Moriyón I
Departamento de Microbiología, Universidad de Navarra, Pamplona, Spain.
J Clin Microbiol. 1994 May;32(5):1159-65. doi: 10.1128/jcm.32.5.1159-1165.1994.
Five serological assays were evaluated for the diagnosis of brucellosis in goats: the rose bengal test (RBT), complement fixation test (CFT), radial immunodiffusion (RID) with Brucella and Yersinia enterocolitica O:9 polysaccharides, counterimmunoelectrophoresis (CIEP) with cytosol, and enzyme-linked immunosorbent assay (ELISA) with polyclonal and protein G conjugates and smooth lipopolysaccharide (S-LPS), native hapten polysaccharide (NH), or cytosol antigens. For optimal sensitivity, RBT had to be used with sera-antigen at a 3:1 dilution. In the RID test, Brucella melitensis biotype 1 NH could not be replaced by Brucella abortus biotype 1 or Y. enterocolitica 0:9 polysaccharides. In the ELISA, S-LPS and NH gave similar results and the protein G conjugate increased the specificity. With the sera from 55 B. melitensis culture-positive goats, the sensitivity was 100% for RBT, CFT (titer > or = 4), and ELISA with S-LPS or NH; 94% for RID; and 93% for CIEP. All tests were negative (100% specific) when testing the sera from 127 brucella-free goats. Larger discrepancies among the results of the serological tests were obtained with sera from goats of areas where brucellosis is endemic. When the sera of 20 young goats vaccinated subcutaneously (10(9) CFU of B. melitensis Rev 1) and bled 6 months later were examined, the specificities were as follows: NH ELISA, 60%; CFT and S-LPS ELISA, 75%; RBT, 80%; CIEP, 90%; and RID, 94%. With the sera from 10 young goats vaccinated conjunctivally (10(9) CFU of B. melitensis Rev 1) all tests were 100% specific 4 months after vaccination. The proportion of goats giving a positive reaction after vaccination decreased faster in RID than in other tests.
对五种血清学检测方法进行了评估，以诊断山羊布鲁氏菌病：玫瑰红试验（RBT）、补体结合试验（CFT）、用布鲁氏菌和小肠结肠炎耶尔森氏菌O:9多糖进行的放射免疫扩散试验（RID）、用细胞溶质进行的对流免疫电泳试验（CIEP），以及用多克隆抗体和蛋白G结合物与光滑脂多糖（S-LPS）、天然半抗原多糖（NH）或细胞溶质抗原进行的酶联免疫吸附试验（ELISA）。为获得最佳灵敏度，RBT必须使用血清与抗原按3:1稀释度混合的样本。在RID试验中，羊种布鲁氏菌生物型1的NH不能被流产布鲁氏菌生物型1或小肠结肠炎耶尔森氏菌O:9多糖替代。在ELISA中，S-LPS和NH得出的结果相似，且蛋白G结合物提高了特异性。用来自55只羊种布鲁氏菌培养阳性山羊的血清进行检测时，RBT、CFT（滴度≥4）以及用S-LPS或NH进行的ELISA的灵敏度均为100%；RID的灵敏度为94%；CIEP 的灵敏度为93%。对127只无布鲁氏菌山羊的血清进行检测时，所有检测均呈阴性（特异性为100%）。在布鲁氏菌病流行地区山羊的血清检测中，血清学检测结果之间的差异更大。对20只皮下接种（10⁹CFU羊种布鲁氏菌Rev 1）并在6个月后采血的幼龄山羊的血清进行检测时，特异性如下：NH ELISA为60%；CFT和S-LPS ELISA为75%；RBT为80%；CIEP为90%；RID为94%。对10只结膜接种（10⁹CFU羊种布鲁氏菌Rev 1）的幼龄山羊的血清进行检测时，接种疫苗4个月后所有检测的特异性均为100%。接种疫苗后呈阳性反应的山羊比例在RID试验中比在其他试验中下降得更快。
