Wagner Carola L, Riess Tanja, Linke Dirk, Eberhardt Christian, Schäfer Andrea, Reutter Sabine, Maggi Ricardo G, Kempf Volkhard A J
Institut für Medizinische Mikrobiologie und Hygiene, University Hospital, Eberhard-Karls-Universität, Elfriede-Aulhorn-Street 6, 72076 Tübingen, Germany.
Int J Med Microbiol. 2008 Oct;298(7-8):579-90. doi: 10.1016/j.ijmm.2008.01.013. Epub 2008 May 22.
Bartonella henselae causes a variety of human diseases (e.g. cat scratch disease and the vasculoproliferative disorders, bacillary angiomatosis and peliosis hepatis). The laboratory diagnosis of B. henselae infections is usually based on the detection of anti-B. henselae antibodies by an indirect immunofluorescence assay (IFA) which, unfortunately, suffers from a significant amount of cross-reactivity and hence is prone to deliver false-positive results. In this pilot study, we evaluated the use of a potential two-step serodiagnosis of B. henselae infections by combining IFA and anti-Bartonella adhesin A (BadA) immunoblotting. Our data revealed that approximately 75% of the IFA-positive sera of patients with a suspected B. henselae infection reacted specifically with BadA but only approximately 25% of the IFA-negative sera of healthy blood donors. Although Yersinia adhesin A (YadA) is structurally closely related to BadA, no cross-reactivity of sera from patients suffering from a Yersinia enterocolitica or Y. pseudotuberculosis infection with BadA was detected in immunoblotting. Unfortunately, recombinantly expressed BadA domains (head, connector, stalk fragment) were not suitable for immunoblotting. Finally, the best resolution for full-length BadA immunoblotting was obtained when whole cell lysates of B. henselae were separated using continuous 4-15% sodium dodecyl sulfate polyacrylamide gels. In summary, our results show that BadA antibodies are detectable in the sera of B. henselae-infected patients and, therefore, this pilot study suggests to include BadA immunoblotting in the laboratory diagnosis of B. henselae infections.
汉赛巴尔通体可引发多种人类疾病（如猫抓病以及血管增生性疾病，杆菌性血管瘤病和肝紫癜）。汉赛巴尔通体感染的实验室诊断通常基于通过间接免疫荧光法（IFA）检测抗汉赛巴尔通体抗体，遗憾的是，该方法存在大量交叉反应，因此容易产生假阳性结果。在这项初步研究中，我们评估了通过结合IFA和抗巴尔通体黏附素A（BadA）免疫印迹对汉赛巴尔通体感染进行潜在两步血清学诊断的应用。我们的数据显示，疑似汉赛巴尔通体感染患者中约75%的IFA阳性血清与BadA发生特异性反应，但健康献血者的IFA阴性血清中只有约25%出现这种情况。尽管耶尔森菌黏附素A（YadA）在结构上与BadA密切相关，但在免疫印迹中未检测到患有小肠结肠炎耶尔森菌或假结核耶尔森菌感染的患者血清与BadA之间存在交叉反应。遗憾的是，重组表达的BadA结构域（头部、连接体、柄片段）不适用于免疫印迹。最后，当使用连续的4-15%十二烷基硫酸钠聚丙烯酰胺凝胶分离汉赛巴尔通体的全细胞裂解物时，获得了全长BadA免疫印迹的最佳分辨率。总之，我们的结果表明在汉赛巴尔通体感染患者的血清中可检测到BadA抗体，因此，这项初步研究建议将BadA免疫印迹纳入汉赛巴尔通体感染的实验室诊断中。
