Cassataro Juliana, Delpino M Victoria, Velikovsky Carlos A, Bruno Laura, Fossati Carlos A, Baldi Pablo C
Instituto de Estudios de la Inmunidad Humoral (IDEHU), Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, 1113 Buenos Aires, Argentina.
Clin Diagn Lab Immunol. 2002 Mar;9(2):366-9. doi: 10.1128/cdli.9.2.366-369.2002.
The diagnostic usefulness of an enzyme-linked immunosorbent assay (ELISA) using a purified recombinant ribosome recycling factor from Brucella melitensis (CP24 antigen) was tested in human and canine infections caused by smooth and rough Brucella species, respectively. Anti-CP24 antibodies were detected in 9 (43%) of 21 consecutive cases of canine brucellosis and in 8 (53%) of 15 dogs followed for 60 days after the diagnosis of acute brucellosis. Among eight patients with acute brucellosis, anti-CP24 antibodies were detected in four in the 10 weeks following diagnosis, but the remaining four were negative during the whole follow-up (22 weeks). The frequency of anti-CP24 antibodies was also low among 24 patients with subacute brucellosis and 23 patients with chronic illness (29 and 26%, respectively). While all patients positive for anti-CP24 antibodies were also positive for antibodies to total cytoplasmic proteins of Brucella (CP), five were negative for antibodies to another cytoplasmic protein, the Brucella lumazine synthase (BLS). When a larger sample of 35 human sera negative for anti-BLS antibodies was assayed, 85.7% were positive for anti-CP24 antibodies, suggesting that the combined measurement of both reactivities could yield a higher sensitivity than any test alone. To test this hypothesis, an ELISA combining both antigens was designed. The percentage of positive results among chronic cases was higher for this assay than for the individual measurement of anti-CP24 or anti-BLS antibodies (83 versus 26 and 65%, respectively) and was closer to the value obtained for anti-CP antibodies (91%). The frequency of anti-CP24 antibodies is low in both canine and human brucellosis. In the latter case, however, an ELISA combining CP24 and BLS is more sensitive than assays measuring anti-CP24 or anti-BLS antibodies separately and almost as sensitive as the ELISA using CP.
利用来自流产布鲁氏菌的纯化重组核糖体循环因子(CP24抗原)的酶联免疫吸附测定(ELISA),分别在由光滑型和粗糙型布鲁氏菌引起的人类和犬类感染中测试其诊断效用。在21例连续的犬布鲁氏菌病病例中有9例(43%)检测到抗CP24抗体,在15只犬急性布鲁氏菌病诊断后随访60天的犬中有8例(53%)检测到抗CP24抗体。在8例急性布鲁氏菌病患者中,诊断后10周内有4例检测到抗CP24抗体,但其余4例在整个随访期间(22周)均为阴性。在24例亚急性布鲁氏菌病患者和23例慢性病患者中,抗CP24抗体的频率也较低(分别为29%和26%)。虽然所有抗CP24抗体阳性的患者布鲁氏菌总细胞质蛋白(CP)抗体也为阳性,但有5例布鲁氏菌核黄素合酶(BLS)这一另一种细胞质蛋白的抗体为阴性。当检测35份抗BLS抗体阴性的人血清大样本时,85.7%抗CP24抗体为阳性,这表明两种反应性的联合检测可能比任何单独检测具有更高的灵敏度。为验证这一假设,设计了一种结合两种抗原的ELISA。该检测在慢性病例中的阳性结果百分比高于抗CP24或抗BLS抗体的单独检测(分别为83%对26%和65%),且更接近抗CP抗体获得的值(91%)。抗CP24抗体在犬类和人类布鲁氏菌病中的频率都较低。然而,在人类病例中,结合CP24和BLS的ELISA比分别检测抗CP24或抗BLS抗体的检测更敏感,几乎与使用CP的ELISA一样敏感。