Culture of Bartonella quintana and Bartonella henselae from Human Samples: a 5-Year Experience (1993 to 1998)

Abstract

Bartonella quintana and Bartonella henselaeare 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 ofBartonella 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 ofB. 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 ofBartonella 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 ofB. 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.

Serological evidence of Bartonella vinsonii lymphadenopathies in a child bitten by a dog

“…Unfortunately, only one blood sample was available for this patient, which remains culture and PCR negative. However, it is well known that the sensitivity of culture is low when compared with that of PCR-based detection methods, even in lymph nodes of CSD [4]. The current case reinforces the hypothesis that any Bartonella may cause human infections, including lymphadenopathy, in patients exposed to the bite of an infected animal.…”

Prevalence of Bartonella henselae antibodies in serum of cats with and without clinical signs of central nervous system disease

“…It is now apparent that some humans with suspected B henselae associated illness were actually infected by B koehlerae (Avidor et al 2004). In the future, prospective studies evaluating for potential disease associations with Bartonella species should use techniques like PCR assay or culture with subsequent genotyping to allow for more stringent organism identification (La Scola and Raoult 1999, Sander et al 1999, Fenollar and Raoult 2004.…”

Modern clinical microbiology: new challenges and solutions

“…A third approach (often occurring by accident) is to extend the incubation time of fastidious extra cellular pathogens, such as the extracellular forms of the facultative intracellular Bartonella spp. 25 , and of intracellular pathogens, such as Bartonella spp., T. whipplei and mycobacteria [25][26][27] . A fourth approach is the use of cell culture, especially the shell vial assay.…”