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. 1997 Mar;35(3):544–547. doi: 10.1128/jcm.35.3.544-547.1997

Isolation of Bartonella (Rochalimaea) henselae: effects of methods of blood collection and handling.

S A Brenner 1, J A Rooney 1, P Manzewitsch 1, R L Regnery 1
PMCID: PMC229623  PMID: 9041385

Abstract

Bartonella (Rochalimaea) henselae causes cat-scratch disease, bacillary angiomatosis, peliosis hepatis, and fever in humans. B. henselae can be difficult to culture axenically, and as many as 5 weeks may be required before colonies are visible. We compared how different methods of blood collection and handling affect isolation of this pathogen. Blood specimens from B. henselae-infected cats were collected in both EDTA and Isolator blood-lysis tubes and were subsequently plated onto rabbit blood-brain heart infusion agar by using three different schedules: plating immediately, plating after 24 h at 25 degrees C, and plating after 26 days at -65 degrees C. Colonies were counted 14 and 35 days after plating. Blood collected in tubes containing EDTA, frozen at -65 degrees C, and then plated on blood agar yielded a median of 60,000 CFU/ml, compared with 25,333 CFU/ml after collection in the Isolator tubes (P < 0.01). Frozen blood yielded the largest number of B. henselae colonies for any of the schedules tested. These results support previous observations that the Isolator system is more sensitive than tubes containing EDTA for isolation of B. henselae and suggest that, for cat blood, collection in tubes containing EDTA and subsequent freezing may further improve the sensitivity of detection of B. henselae.

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Selected References

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  1. Anderson B., Sims K., Regnery R., Robinson L., Schmidt M. J., Goral S., Hager C., Edwards K. Detection of Rochalimaea henselae DNA in specimens from cat scratch disease patients by PCR. J Clin Microbiol. 1994 Apr;32(4):942–948. doi: 10.1128/jcm.32.4.942-948.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Batterman H. J., Peek J. A., Loutit J. S., Falkow S., Tompkins L. S. Bartonella henselae and Bartonella quintana adherence to and entry into cultured human epithelial cells. Infect Immun. 1995 Nov;63(11):4553–4556. doi: 10.1128/iai.63.11.4553-4556.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benson L. A., Kar S., McLaughlin G., Ihler G. M. Entry of Bartonella bacilliformis into erythrocytes. Infect Immun. 1986 Nov;54(2):347–353. doi: 10.1128/iai.54.2.347-353.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brannon P., Kiehn T. E. Large-scale clinical comparison of the lysis-centrifugation and radiometric systems for blood culture. J Clin Microbiol. 1985 Dec;22(6):951–954. doi: 10.1128/jcm.22.6.951-954.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cashman J. S., Boshard R., Matsen J. M. Viability of organisms held in the isolator blood culture system for 15 h and their rapid detection by acridine orange staining. J Clin Microbiol. 1983 Sep;18(3):709–712. doi: 10.1128/jcm.18.3.709-712.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Garcia F. U., Wojta J., Hoover R. L. Interactions between live Bartonella bacilliformis and endothelial cells. J Infect Dis. 1992 Jun;165(6):1138–1141. doi: 10.1093/infdis/165.6.1138. [DOI] [PubMed] [Google Scholar]
  7. Henry N. K., McLimans C. A., Wright A. J., Thompson R. L., Wilson W. R., Washington J. A., 2nd Microbiological and clinical evaluation of the isolator lysis-centrifugation blood culture tube. J Clin Microbiol. 1983 May;17(5):864–869. doi: 10.1128/jcm.17.5.864-869.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hill E. M., Raji A., Valenzuela M. S., Garcia F., Hoover R. Adhesion to and invasion of cultured human cells by Bartonella bacilliformis. Infect Immun. 1992 Oct;60(10):4051–4058. doi: 10.1128/iai.60.10.4051-4058.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kiehn T. E., Wong B., Edwards F. F., Armstrong D. Comparative recovery of bacteria and yeasts from lysis-centrifugation and a conventional blood culture system. J Clin Microbiol. 1983 Aug;18(2):300–304. doi: 10.1128/jcm.18.2.300-304.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Koehler J. E., Glaser C. A., Tappero J. W. Rochalimaea henselae infection. A new zoonosis with the domestic cat as reservoir. JAMA. 1994 Feb 16;271(7):531–535. doi: 10.1001/jama.271.7.531. [DOI] [PubMed] [Google Scholar]
  11. Kordick D. L., Breitschwerdt E. B. Intraerythrocytic presence of Bartonella henselae. J Clin Microbiol. 1995 Jun;33(6):1655–1656. doi: 10.1128/jcm.33.6.1655-1656.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lucey D., Dolan M. J., Moss C. W., Garcia M., Hollis D. G., Wegner S., Morgan G., Almeida R., Leong D., Greisen K. S. Relapsing illness due to Rochalimaea henselae in immunocompetent hosts: implication for therapy and new epidemiological associations. Clin Infect Dis. 1992 Mar;14(3):683–688. doi: 10.1093/clinids/14.3.683. [DOI] [PubMed] [Google Scholar]
  13. Mernaugh G., Ihler G. M. Deformation factor: an extracellular protein synthesized by Bartonella bacilliformis that deforms erythrocyte membranes. Infect Immun. 1992 Mar;60(3):937–943. doi: 10.1128/iai.60.3.937-943.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Noah D. L., Bresee J. S., Gorensek M. J., Rooney J. A., Cresanta J. L., Regnery R. L., Wong J., del Toro J., Olson J. G., Childs J. E. Cluster of five children with acute encephalopathy associated with cat-scratch disease in south Florida. Pediatr Infect Dis J. 1995 Oct;14(10):866–869. doi: 10.1097/00006454-199510000-00009. [DOI] [PubMed] [Google Scholar]
  15. Norman A. F., Regnery R., Jameson P., Greene C., Krause D. C. Differentiation of Bartonella-like isolates at the species level by PCR-restriction fragment length polymorphism in the citrate synthase gene. J Clin Microbiol. 1995 Jul;33(7):1797–1803. doi: 10.1128/jcm.33.7.1797-1803.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Regnery R. L., Anderson B. E., Clarridge J. E., 3rd, Rodriguez-Barradas M. C., Jones D. C., Carr J. H. Characterization of a novel Rochalimaea species, R. henselae sp. nov., isolated from blood of a febrile, human immunodeficiency virus-positive patient. J Clin Microbiol. 1992 Feb;30(2):265–274. doi: 10.1128/jcm.30.2.265-274.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Regnery R. L., Olson J. G., Perkins B. A., Bibb W. Serological response to "Rochalimaea henselae" antigen in suspected cat-scratch disease. Lancet. 1992 Jun 13;339(8807):1443–1445. doi: 10.1016/0140-6736(92)92032-b. [DOI] [PubMed] [Google Scholar]
  18. Regnery R., Martin M., Olson J. Naturally occurring "Rochalimaea henselae" infection in domestic cat. Lancet. 1992 Aug 29;340(8818):557–558. doi: 10.1016/0140-6736(92)91760-6. [DOI] [PubMed] [Google Scholar]
  19. Regnery R., Tappero J. Unraveling mysteries associated with cat-scratch disease, bacillary angiomatosis, and related syndromes. Emerg Infect Dis. 1995 Jan-Mar;1(1):16–21. doi: 10.3201/eid0101.950103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Relman D. A., Loutit J. S., Schmidt T. M., Falkow S., Tompkins L. S. The agent of bacillary angiomatosis. An approach to the identification of uncultured pathogens. N Engl J Med. 1990 Dec 6;323(23):1573–1580. doi: 10.1056/NEJM199012063232301. [DOI] [PubMed] [Google Scholar]
  21. Rodriguez-Barradas M. C., Bandres J. C., Hamill R. J., Trial J., Clarridge J. E., 3rd, Baughn R. E., Rossen R. D. In vitro evaluation of the role of humoral immunity against Bartonella henselae. Infect Immun. 1995 Jun;63(6):2367–2370. doi: 10.1128/iai.63.6.2367-2370.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Slater L. N., Welch D. F., Hensel D., Coody D. W. A newly recognized fastidious gram-negative pathogen as a cause of fever and bacteremia. N Engl J Med. 1990 Dec 6;323(23):1587–1593. doi: 10.1056/NEJM199012063232303. [DOI] [PubMed] [Google Scholar]
  23. Tompkins L. S. Rochalimaea infections. Are they zoonoses? JAMA. 1994 Feb 16;271(7):553–554. [PubMed] [Google Scholar]
  24. Welch D. F., Hensel D. M., Pickett D. A., San Joaquin V. H., Robinson A., Slater L. N. Bacteremia due to Rochalimaea henselae in a child: practical identification of isolates in the clinical laboratory. J Clin Microbiol. 1993 Sep;31(9):2381–2386. doi: 10.1128/jcm.31.9.2381-2386.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wong M. T., Thornton D. C., Kennedy R. C., Dolan M. J. A chemically defined liquid medium that supports primary isolation of Rochalimaea (Bartonella) henselae from blood and tissue specimens. J Clin Microbiol. 1995 Mar;33(3):742–744. doi: 10.1128/jcm.33.3.742-744.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]

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