Detection of Bartonella (Rochalimaea) quintana by routine acridine orange staining of broth blood cultures

Larson, A.; Dougherty, Molly J.; Nowowiejski, D J; Welch, David; Matar, Ghassan M.; Swaminathan, Bala; Coyle, Marie B.

doi:10.1128/jcm.32.6.1492-1496.1994

Cited by 55 publications

(24 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…1 for Bartonella species and Fig. 2 16,107, and 260 bp for C. ochracea. Experimental results differed from those predicted in that bands with a molecular size of Ͻ200 bp were not easily detected in agarose gel (Fig.…”

Section: Resultsmentioning

confidence: 97%

“…Bartonellae were isolated mainly from blood, although a few isolates were recovered from other tissues, including skin and bone (14). A large series of 34 isolates was recently detected in the blood from 10 alcoholic patients by systematic acridine orange staining of the bottle supernatant (16,35) after the automatic infrared detection system had identified the specimens as negative blood samples. B. quintana and B. henselae were isolated from the blood of immunocompromised patients with relapsing bacteremic fever, and both species, as well as B. elizabethae, produced infective endocarditis.…”

Section: Discussionmentioning

confidence: 99%

“…and incubated for 72 h at 35ЊC. An aliquot was then stained with acridine orange as previously reported (16) and was also Gram stained. Total DNA was extracted from 200 ml of broth supernatant from each bottle by using the QIAamp_ Blood kit (Qiagen, Inc., Chatsworth, Calif.) according to the recommendations of the supplier and was submitted to two cycles of PCR as described above and RFLP analysis as described above.…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Identification of Bartonella (Rochalimaea) species among fastidious gram-negative bacteria on the basis of the partial sequence of the citrate-synthase gene

et al. 1995

View full text Add to dashboard Cite

The bacterial genus Bartonella (Rochalimaea) includes emerging human pathogens with five recognized species. These are fastidious gram-negative bacteria, exhibiting few phenotypic characteristics and whose identification relies upon serotyping, cellular fatty acid analysis, and molecular typing. Most of the isolates have been recovered from the blood of patients, and three of the four pathogenic Bartonella species are associated with infectious endocarditis. We performed PCR-restriction fragment length polymorphism (RFLP) analysis of the blood culture bottle supernatant for the routine identification of Bartonella species among fastidious gram-negative bacteria. The amplification of the citrate-synthase gene with primers previously reported (R. L. Regnery, C. L. Spruill, and B. D. Plikaytis, J. Bacteriol. 173:1576-1589, 1991) yielded a 379-bp product from Bartonella species and a 382-bp product for Capnocytophaga ochracea but no product from any of the other 15 genotypically or phenotypically related species tested. We determined the sequences of the citrate-synthase gene-amplified products for Bartonella species and C. ochracea in order to predict the optimal restriction enzyme to be used in RFLP analysis. TaqI and AciI allowed identification of Bartonella species and C. ochracea. We propose that acridine orange and Gram staining, followed by PCR-RFLP analysis of the blood bottle supernatant, be included in the examination of blood samples from patients with suspected infectious endocarditis.

show abstract

Section: Resultsmentioning

confidence: 97%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Identification of Bartonella (Rochalimaea) species among fastidious gram-negative bacteria on the basis of the partial sequence of the citrate-synthase gene

et al. 1995

View full text Add to dashboard Cite

show abstract

“…More recently, both B. henselae and B. quintana have been associated with bacillary angiomatosis, bacteremia, and prolonged febrile illness (8,9,11,14,15,16,20,21,26,27). Isolation of the organism is usually from blood, but isolation can also be achieved from tissue, although with more difficulty (1,5,9,10,13). Although it has been proposed that Afipia felis is the etiologic agent of cat scratch disease, more recent studies by PCR, culture, and serology strongly indicate that B. henselae is the infecting species (1,5,22).…”

mentioning

confidence: 99%

Strategy to detect and identify Bartonella species in routine clinical laboratory yields Bartonella henselae from human immunodeficiency virus-positive patient and unique Bartonella strain from his cat

et al. 1995

View full text Add to dashboard Cite

We wished to develop a cost-effective, rapid strategy to detect and identify Bartonella species in the clinical laboratory and to determine the prevalence of Bartonella infection in the Houston veteran population. Bartonella colonies were identified by colony morphology, Gram stain, RapID ANA, repetitive extragenic palindromic-PCR (REP-PCR) and whole-cell fatty acid (CFA) analysis, and these methods were compared for their usefulness. A new test order for ''Rochalimaea culture'' (the genus Bartonella was previously known as the genus Rochalimaea) was instituted, and in addition, all blood specimens submitted for fungal culture (obtained in an isolator tube) were processed for Bartonella culture. Over a 16-month period we isolated Bartonella henselae from only 0.4% (2 of 533) of total cultures but from 1% (2 of 204) of human immunodeficiency virus-positive patients. After sufficient growth, identification of the Bartonella isolates to the species level could be obtained in 2 days. The REP-PCR allowed discrimination of all known species, whereas CFA analysis distinguished all except B. henselae and Bartonella quintana. The RapID ANA results failed to differentiate between B. henselae and B. quintana, and results for other species differed by only one or two tests. Blood obtained from a kitten which had been introduced into the household of one patient 2 months before the onset of fever yielded a Bartonella strain which was shown to be different from the strain from the patient and distinct from other Bartonella species by a combination of REP-PCR, CFA, and growth characteristics. Subsequent analysis of the citrate synthase gene sequence showed only an 86% similarity with any of the other known Bartonella species, suggesting that this isolate represents a distinct, previously uncharacterized species of Bartonella.

show abstract

“…Other fastidious organisms isolated from lower respiratory tract infections in HIV-infected patients and capable of dissemination include Mycobacterium malmoense (17) and nutritionally aberrant Mycobacterium tuberculosis (5). Recent recommendations for modifying blood culture protocols to enhance recovery of these fastidious organisms include extended incubation times (8), lowered incubation temperatures (4,6), supplemented media (2,6), and terminal examination of broth media by subculture (16) or acridine orange stain (8).…”

mentioning

confidence: 99%

Evaluation of an extended blood culture protocol to isolate fastidious organisms from patients with AIDS

et al. 1996

Self Cite

View full text Add to dashboard Cite

Recent reports of fastidious pathogens suggest the need for special blood cultures for immunocompromised patients. Blood cultures from 45 human immunodeficiency virus (HIV)-infected patients with unexplained fever (>38.0؇C) and CD4 counts of <125 cells per mm 3 were collected into a vacuum tube with sodium polyanetholsulfonate, an Isolator tube, and BACTEC aerobic and anaerobic bottles. Blood from the sodium polyanethosulfonate tube was inoculated into BACTEC 13A bottles, which were read weekly for 16 weeks. Isolator sediment was divided among eight agar media, including four sheep blood agar media: chocolate agar, brain heart infusion blood agar, heart infusion blood agar, and brucella blood agar. Other agar plates included Sabouraud's, buffered charcoal-yeast extract, Middlebrook 7H11 (M7H11) with hemoglobin, and M7H11 with mycobactin J. Incubation conditions included air and CO 2 -enriched aerobic, microaerophilic, and anaerobic atmospheres. Aerobic BACTEC broths received an acridine orange stain on day 8 and were subcultured at 2, 4, and 8 weeks. Anaerobic BACTEC bottles were subcultured at 4 weeks. All solid media, including subcultures, were incubated for 8 weeks, providing a total of 16 weeks of incubation for each specimen. Clinically significant isolates included eight Mycobacterium avium complex isolates and one each of Bartonella henselae, Bartonella quintana, Shigella flexneri, Klebsiella oxytoca, and Cryptococcus neoformans. All isolates were detected with commercially available media and, with the exception of Bartonella spp., were recovered within incubation times routinely used in most clinical laboratories.

show abstract

Detection of Bartonella (Rochalimaea) quintana by routine acridine orange staining of broth blood cultures

Cited by 55 publications

References 20 publications

Identification of Bartonella (Rochalimaea) species among fastidious gram-negative bacteria on the basis of the partial sequence of the citrate-synthase gene

Identification of Bartonella (Rochalimaea) species among fastidious gram-negative bacteria on the basis of the partial sequence of the citrate-synthase gene

Strategy to detect and identify Bartonella species in routine clinical laboratory yields Bartonella henselae from human immunodeficiency virus-positive patient and unique Bartonella strain from his cat

Evaluation of an extended blood culture protocol to isolate fastidious organisms from patients with AIDS

Contact Info

Product

Resources

About