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Conservation of the 17-Kilodalton Antigen Gene within the Genus Bartonella

机译:Bartonella属中的17-Kilodalton抗原基因的保守性

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The 17-kDa antigen of Bartonella henselae has previously been shown to elicit a strong humoral immune response in patients with cat scratch disease (CSD) and to be useful in screening human serum samples for CSD. In this study, PCR amplification of genes homologous to the 17-kDa antigen gene of B. henselae was performed using genomic DNAs from several species ofBartonella, including the currently recognized human pathogens. Amplicons of similar size were demonstrated using the following chromosomal DNA templates: B. henselae (two strains), B. quintana (two strains), B. elizabethae, B. clarridgeiae, B. vinsoniisubsp. vinsonii, and B. vinsonii subsp.berkhoffii. No evidence of a B. bacilliformishomolog of the 17-kDa antigen gene was obtained using multiple primer pairs. DNA sequencing revealed open reading frames capable of coding for proteins with sizes similar to that of the 17-kDa antigen ofB. henselae in all of the amplicons; however, extensive sequence divergence across the genus was noted. Cloning of the amplified products into pUC19 resulted in recombinants that directed synthesis of homologs of the 17-kDa protein. Immunoblot analysis using human sera from CSD cases demonstrated very little cross-reactivity among different species for this protein. In contrast, immunoblots using rabbit serum raised to the recombinant B. henselaeantigen showed extensive cross-reactivity with the proteins of otherBartonella species. The data suggest that the use of the 17-kDa antigen as a serologic reagent may allow the development of more specific diagnostic assays. Furthermore, the nucleotide sequences from the various versions of the 17-kDa antigen gene should be useful for rapid identification of Bartonella at the species level.
机译:先前已证明 Bartonella henselae 的17-kDa抗原在猫抓痕病(CSD)患者中引起强烈的体液免疫反应,可用于筛查CSD的人血清样品。在这项研究中,PCR扩增了与 B的17-kDa抗原基因同源的基因。使用来自 Bartonella 几种物种的基因组DNA进行henselae ,包括目前公认的人类病原体。使用以下染色体DNA模板证明了类似大小的扩增子: B。 henselae (两种菌株), B。金塔纳(两种菌株), B。 elizabethae B。 clarridgeiae B。 vinsonii subsp。 vinsonii B。 vinsonii 亚种 berkhoffii 。没有 B的证据。使用多个引物对获得了17-kDa抗原基因的芽孢杆菌同源性。 DNA测序揭示了开放阅读框,其能够编码大小类似于em B的17-kDa抗原的蛋白质。 henselae 在所有扩增子中;然而,注意到该属广泛的序列差异。将扩增的产物克隆到pUC19中产生了指导17-kDa蛋白同源物合成的重组体。使用来自CSD病例的人血清进行的免疫印迹分析表明,不同物种之间对该蛋白的交叉反应极少。相反,使用兔血清的免疫印迹可产生重组B。 henselae 抗原与其他 Bartonella 物种的蛋白质具有广泛的交叉反应性。数据表明,将17-kDa抗原用作血清学试剂可以开发更特异性的诊断方法。此外,来自17-kDa抗原基因各种版本的核苷酸序列对于在物种水平上快速鉴定Bartonella 很有用。

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