La Scola B, Raoult D
Unité des Rickettsies, CNRS UPRESA 6020, Faculté de Médecine, 13385 Marseille Cedex 05, France.
J Clin Microbiol. 1999 Jun;37(6):1899-905. doi: 10.1128/JCM.37.6.1899-1905.1999.
Bartonella quintana and Bartonella henselae are fastidious gram-negative bacteria responsible for bacillary angiomatosis, trench fever, cat scratch disease, and endocarditis. During a 5-year period, we received 2,043 samples for culture of Bartonella sp. We found Bartonella sp. to be the etiologic agent in 38 cases of endocarditis, 78 cases of cat scratch disease, 16 cases of bacteremia in homeless people, and 7 cases of bacillary angiomatosis. We correlated the results of positive cultures with the clinical form of the disease, type of sample, culture procedure, PCR-based genomic detection, and antibody determination. Seventy-two isolates of B. quintana and nine isolates of B. henselae from 43 patients were obtained. Sixty-three of the B. quintana isolates and two of the B. henselae isolates, obtained from patients with no prior antibiotic therapy, were stably subcultured. The sensitivity of culture was low when compared with that of PCR-based detection methods in valves of patients with endocarditis (44 and 81%, respectively), skin biopsy samples of patients with bacillary angiomatosis (43 and 100%, respectively), and lymph nodes of cat scratch disease (13 and 30%, respectively). Serological diagnosis was also more sensitive in cases of endocarditis (97%) and cat scratch disease (90%). Among endocarditis patients, the sensitivity of the shell vial culture assay was 28% when inoculated with blood samples and 44% when inoculated with valvular biopsy samples, and the sensitivity of both was significantly higher than that of culture on agar (5% for blood [P = 0.045] and 4% for valve biopsy samples [P < 0.0005]). The most efficient culture procedure was the subculture of blood culture broth into shell vials (sensitivity, 71%). For patients with endocarditis, previous antibiotic therapy significantly affected results of blood culture; no patient who had been administered antibiotics yielded a positive blood culture, whereas 80% of patients with no previous antibiotic therapy yielded positive blood cultures (P = 0.0006). Previous antibiotic therapy did not, however, prevent isolation of Bartonella sp. from cardiac valves but did prevent the establishment of strains, as none of the 15 isolates from treated patients could be successfully subcultured. For the diagnosis of B. quintana bacteremia in homeless people, the efficiency of systematic subculture of blood culture broth onto agar was higher than that of direct blood plating (respective sensitivities, 98 and 10% [P < 10(-7)]). Nevertheless, both procedures are complementary, since when used together their sensitivity reached 100%. All homeless people with positive blood cultures had negative serology. The isolation rate of B. henselae from PCR-positive lymph nodes, in patients with cat scratch disease, was significantly lower than that from valves of endocarditis patients and skin biopsy samples from bacillary angiomatosis patients (13 and 33%, respectively [P = 0.084]). In cases of bacillary angiomatosis for which an agent was identified to species level, the isolation rate of B. henselae was lower than the isolation rate of B. quintana (28 and 64%, respectively [P = 0.003]). If culture is to be considered an efficient tool for the diagnosis of several Bartonella-related diseases, methodologies need to be improved, notably for the recovery of B. henselae from lymph nodes of patients with cat scratch disease.
五日热巴尔通体和汉赛巴尔通体是苛养革兰氏阴性菌,可引起杆菌性血管瘤病、战壕热、猫抓病和心内膜炎。在5年期间,我们收到了2043份用于培养巴尔通体属细菌的样本。我们发现巴尔通体属细菌是38例心内膜炎、78例猫抓病、16例无家可归者菌血症和7例杆菌性血管瘤病的病原体。我们将阳性培养结果与疾病的临床形式、样本类型、培养程序、基于PCR的基因组检测和抗体测定进行了关联。从43例患者中获得了72株五日热巴尔通体分离株和9株汉赛巴尔通体分离株。从无先前抗生素治疗的患者中获得的63株五日热巴尔通体分离株和2株汉赛巴尔通体分离株能够稳定传代培养。与基于PCR的检测方法相比,在心内膜炎患者的瓣膜(分别为44%和81%)、杆菌性血管瘤病患者的皮肤活检样本(分别为43%和100%)以及猫抓病患者的淋巴结(分别为13%和30%)中,培养的敏感性较低。血清学诊断在心内膜炎(97%)和猫抓病(90%)病例中也更敏感。在心内膜炎患者中,用血液样本接种时,壳瓶培养试验的敏感性为28%,用瓣膜活检样本接种时为44%,两者的敏感性均显著高于琼脂培养(血液为5%[P = 0.045],瓣膜活检样本为4%[P < 0.0005])。最有效的培养程序是将血培养肉汤传代接种到壳瓶中(敏感性为71%)。对于心内膜炎患者,先前的抗生素治疗显著影响血培养结果;接受过抗生素治疗的患者中没有一人血培养呈阳性,而未接受过抗生素治疗的患者中有80%血培养呈阳性(P = 0.0006)。然而,先前的抗生素治疗并未阻止从心脏瓣膜中分离出巴尔通体属细菌,但确实阻止了菌株的建立,因为从接受治疗的患者中分离出的15株菌株均无法成功传代培养。对于诊断无家可归者的五日热巴尔通体菌血症,将血培养肉汤系统传代接种到琼脂上的效率高于直接血平板接种(敏感性分别为98%和10%[P < 10⁻⁷])。然而,这两种方法是互补的,因为一起使用时它们的敏感性达到了100%。所有血培养呈阳性的无家可归者血清学均为阴性。在猫抓病患者中,从PCR阳性淋巴结中分离出汉赛巴尔通体的比率显著低于从心内膜炎患者的瓣膜和杆菌性血管瘤病患者的皮肤活检样本中分离出的比率(分别为13%和33%[P = 0.084])。在确定病原体到种水平的杆菌性血管瘤病病例中,汉赛巴尔通体的分离率低于五日热巴尔通体的分离率(分别为28%和64%[P = 0.003])。如果要将培养视为诊断几种巴尔通体相关疾病的有效工具,就需要改进方法,特别是从猫抓病患者的淋巴结中回收汉赛巴尔通体的方法。