Houpikian Pierre, Raoult Didier
Unité des Rickettsies, CNRS-UPRES-A 6020, Faculté de Médecine de Marseille, 13385 Marseille cedex, France.
Clin Diagn Lab Immunol. 2003 Jan;10(1):95-102. doi: 10.1128/cdli.10.1.95-102.2003.
To differentiate infectious endocarditis (IE) from other Bartonella infections and to identify infecting Bartonella bacteria at the species level on a serological basis, we used Western immunoblotting to test sera from 51 patients with Bartonella IE (of which 27 had previously benefited from species identification by molecular techniques), 11 patients with chronic Bartonella quintana bacteremia, and 10 patients with cat scratch disease. Patients with IE were Western blot positive in 49 of 51 cases, and significant cross-reactivity with three heterologous Bartonella antigens was found in 45 of 49 cases. Sera from bacteremic patients did not react with more than one heterologous antigen, and sera from patients with cat scratch disease gave negative results. Sera reacted only with B. henselae in four cases of IE, including one with a positive PCR result for valve tissue. Western blot and cross-adsorption performed on serum samples from patients with IE (the identity of the causative species having been determined by PCR) were demonstrated to identify efficiently the causative species in all cases. When applied to patients diagnosed on the basis of serological tests only, this technique allowed identification of the causative species in 20 of 22 cases. The results were in accordance with epidemiological features. Six reactive bands of B. quintana (of molecular sizes from 10 to 83 kDa) demonstrated significant association with sera from patients with B. quintana endocarditis. Overall, Western blotting and cross-adsorption made it possible to identify the causative species in 49 of 51 (96%) IE cases.
为了将感染性心内膜炎(IE)与其他巴尔通体感染区分开来,并在血清学基础上在种属水平鉴定感染的巴尔通体细菌,我们使用蛋白质免疫印迹法检测了51例巴尔通体性IE患者(其中27例先前已通过分子技术进行了种属鉴定)、11例慢性五日热巴尔通体菌血症患者和10例猫抓病患者的血清。51例IE患者中有49例蛋白质免疫印迹呈阳性,49例中的45例发现与三种异源巴尔通体抗原有显著交叉反应。菌血症患者的血清与不超过一种异源抗原发生反应,猫抓病患者的血清检测结果为阴性。在4例IE患者中,血清仅与汉赛巴尔通体发生反应,其中1例瓣膜组织PCR结果为阳性。对IE患者(致病菌种已通过PCR确定)的血清样本进行蛋白质免疫印迹和交叉吸附,结果表明在所有病例中均能有效鉴定致病菌种。当应用于仅根据血清学检测诊断的患者时,该技术在22例中的20例中鉴定出了致病菌种。结果与流行病学特征相符。五日热巴尔通体的6条反应带(分子大小为10至83 kDa)与五日热巴尔通体心内膜炎患者的血清有显著相关性。总体而言,蛋白质免疫印迹和交叉吸附使得在51例IE病例中的49例(96%)中鉴定出致病菌种成为可能。