1887

Abstract

A detection system for DNA in blood samples based on the PCR was developed and evaluated in A/J mice with experimentally induced pneumonia. Primers were designed to amplify a 106 bp DNA fragment of the 16S rRNA gene specific to species. The PCR system could detect clinically relevant species including , , , , , and . The sensitivity of the PCR system was 20 fg extracted DNA. In the mouse model, the blood PCR was compared with results obtained by PCR on bronchoalveolar lavage fluid (BALF) samples, cultures of blood and BALF and detection of urinary antigen. Blood PCR was positive until 8 days after infection, while BALF PCR became negative on day 4. These results indicate that PCR using blood samples may be a useful, convenient and non-invasive method for the diagnosis of pneumonia.

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2003-04-01
2019-10-18
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References

  1. Aebischer, C. C., Matter, L., Gaia, V. & Aebi, C. ( 1999;). Diagnosis by polymerase chain reaction of pneumonia caused by Legionella pneumophila in an immunocompetent child. Infection 27, 280–282.[CrossRef]
    [Google Scholar]
  2. Benson, R. F., Tang, P. W. & Fields, B. S. ( 2000;). Evaluation of the Binax and Biotest urinary antigen kits for detection of Legionnaires’ disease due to multiple serogroups and species of Legionella. J Clin Microbiol 38, 2763–2765.
    [Google Scholar]
  3. Breiman, R. F. & Butler, J. C. ( 1998;). Legionnaires’ disease: clinical, epidemiological, and public health perspectives. Semin Respir Infect 13, 84–89.
    [Google Scholar]
  4. Brieland, J., Freeman, P., Kunkel, R., Chrisp, C., Hurley, M., Fantone, J. & Engleberg, C. ( 1994;). Replicative Legionella pneumophila lung infection in intratracheally inoculated A/J mice. A murine model of human Legionnaires’ disease. Am J Pathol 145, 1537–1546.
    [Google Scholar]
  5. Clark, N. C., Cooksey, R. C., Hill, B. C., Swenson, J. M. & Tenover, F. C. ( 1993;). Characterization of glycopeptide-resistant enterococci from US hospitals. Antimicrob Agents Chemother 37, 2311–2317.[CrossRef]
    [Google Scholar]
  6. Cloud, J. L., Carroll, K. C., Pixton, P., Erali, M. & Hillyard, D. R. ( 2000;). Detection of Legionella species in respiratory specimens using PCR with sequencing confirmation. J Clin Microbiol 38, 1709–1712.
    [Google Scholar]
  7. Dominguez, J. A., Gali, N., Pedroso, P., Fargas, A., Padilla, E., Manterola, J. M. & Matas, L. ( 1998;). Comparison of the Binax Legionella urinary antigen enzyme immunoassay (EIA) with the Biotest Legionella Urine antigen EIA for detection of Legionella antigen in both concentrated and nonconcentrated urine samples. J Clin Microbiol 36, 2718–2722.
    [Google Scholar]
  8. El-Solh, A. A., Sikka, P., Ramadan, F. & Davies, J. ( 2001;). Etiology of severe pneumonia in the very elderly. Am J Respir Crit Care Med 163, 645–651.[CrossRef]
    [Google Scholar]
  9. Engleberg, N. C., Carter, C., Weber, D. R., Cianciotto, N. P. & Eisenstein, B. I. ( 1989;). DNA sequence of mip, a Legionella pneumophila gene associated with macrophage infectivity. Infect Immun 57, 1263–1270.
    [Google Scholar]
  10. Harrison, T. G. & Taylor, A. G. ( 1988;). Timing of seroconversion in Legionnaires’ disease. Lancet ii, 795.
    [Google Scholar]
  11. Helbig, J. H., Engelstadter, T., Maiwald, M., Uldum, S. A., Witzleb, P. C. & Luck, P. C. ( 1999;). Diagnostic relevance of the detection of Legionella DNA in urine samples by the polymerase chain reaction. Eur J Clin Microbiol Infect Dis 18, 716–722.[CrossRef]
    [Google Scholar]
  12. Hirakata, Y., Hagiwara, S., Kawaguchi, K., Ishii, Y., Sugiyama, Y. & Kitamura, S. ( 1996;). Two coincidental isolated cases of Legionnaires’ disease diagnosed by PCR and cured by combination therapy with clarithromycin and rifampicin. J Infect Chemother 2, 187–193.[CrossRef]
    [Google Scholar]
  13. Iwamoto, M., Koga, H., Kohno, S., Kaku, M. & Hara, K. ( 1994;). Detection of Legionella species by polymerase chain reaction. Infect Dis 7, 99–103.
    [Google Scholar]
  14. Jaulhac, B., Reyrolle, M., Sodahlon, Y. K., Jarraud, S., Kubina, M., Monteil, H., Piemont, Y. & Etienne, J. ( 1998;). Comparison of sample preparation methods for detection of Legionella pneumophila in culture-positive bronchoalveolar lavage fluids by PCR. J Clin Microbiol 36, 2120–2122.
    [Google Scholar]
  15. Jonas, D., Rosenbaum, A., Weyrich, S. & Bhakdi, S. ( 1995;). Enzyme-linked immunoassay for detection of PCR-amplified DNA of legionellae in bronchoalveolar fluid. J Clin Microbiol 33, 1247–1252.
    [Google Scholar]
  16. Lindsay, D. S., Abraham, W. H. & Fallon, R. J. ( 1994;). Detection of mip gene by PCR for diagnosis of Legionnaires’ disease. J Clin Microbiol 32, 3068–3069.
    [Google Scholar]
  17. Lo Presti, F., Riffard, S., Jarraud, S., Le Gallou, F., Richet, H., Vandenesch, F. & Etienne, J. ( 2000;). Isolation of Legionella oakridgensis from two patients with pleural effusion living in the same geographical area. J Clin Microbiol 38, 3128–3130.
    [Google Scholar]
  18. Maiwald, M., Schill, M., Stockinger, C., Helbig, J. H., Luck, P. C., Witzleb, W. & Sonntag, H.-G. ( 1995;). Detection of Legionella DNA in human and guinea pig urine samples by the polymerase chain reaction. Eur J Clin Microbiol Infect Dis 14, 25–33.[CrossRef]
    [Google Scholar]
  19. Marston, B. J., Plouffe, J. F., File, T. M., Jr, Hackman, B. A., Salstrom, S.-J., Lipman, H. B., Kolczak, M. S. & Breiman, R. F. ( 1997;). Incidence of community-acquired pneumonia requiring hospitalization. Results of a population-based active surveillance study in Ohio. The Community-Based Pneumonia Incidence Study Group. Arch Intern Med 157, 1709–1718.[CrossRef]
    [Google Scholar]
  20. Matsiota-Bernard, P., Pitsouni, E., Legakis, N. & Nauciel, C. ( 1994;). Evaluation of commercial amplification kit for detection of Legionella pneumophila in clinical specimens. J Clin Microbiol 32, 1503–1505.
    [Google Scholar]
  21. Murdoch, D. R. & Chambers, S. T. ( 2000;). Detection of Legionella DNA in peripheral leukocytes, serum, and urine from a patient with pneumonia caused by Legionella dumoffii. Clin Infect Dis 30, 382–383.[CrossRef]
    [Google Scholar]
  22. Murdoch, D. R., Walford, E. J., Jennings, L. C., Light, G. J., Schousboe, M. I., Chereshsky, A. Y., Chambers, S. T. & Town, G. I. ( 1996;). Use of the polymerase chain reaction to detect Legionella DNA in urine and serum samples from patients with pneumonia. Clin Infect Dis 23, 475–480.[CrossRef]
    [Google Scholar]
  23. Murdoch, D. R., Jennings, L. C., Light, G. J. & Chambers, S. T. ( 1999;). Detection of Legionella DNA in guinea pig peripheral leukocytes, urine and plasma by the polymerase chain reaction. Eur J Clin Microbiol Infect Dis 18, 445–447.[CrossRef]
    [Google Scholar]
  24. Ratcliff, R. M., Lanser, J. A., Manning, P. A. & Heuzenroeder, M. W. ( 1998;). Sequence-based classification scheme for the genus Legionella targeting the mip gene. J Clin Microbiol 36, 1560–1567.
    [Google Scholar]
  25. Ratcliff, R. M., Lanser, J. A., Heuzenroeder, M. W. & Manning, P. A. (2001). Sequence-based genotype scheme for Legionella. In Legionella, pp. 237–242. Edited by R. Marre, Y. A. Kwaki, C. Bartlett, N. P. Cianciotto, B. S. Fields, M. Frosch, J. Hacker & P. C. Luck. Washington, DC: American Society for Microbiology.
  26. Socan, M., Kese, D. & Marinic-Fiser, N. ( 2000;). Polymerase chain reaction for detection of legionellae DNA in urine samples from patients with community-acquired pneumonia. Folia Microbiol (Praha) 45, 469–472.[CrossRef]
    [Google Scholar]
  27. Sopena, N., Sabria, M., Pedro-Botet, M. L. & 7 other authors ( 1999;). Prospective study of community-acquired pneumonia of bacterial etiology in adults. Eur J Clin Microbiol Infect Dis 18, 852–858.[CrossRef]
    [Google Scholar]
  28. Stout, J. E. & Yu, V. L. ( 1997;). Legionellosis. N Engl J Med 337, 682–687.[CrossRef]
    [Google Scholar]
  29. Waterer, G. W., Baselski, V. S. & Wunderink, R. G. ( 2001;). Legionella and community-acquired pneumonia: a review of current diagnostic tests from a clinician's viewpoint. Am J Med 110, 41–48.[CrossRef]
    [Google Scholar]
  30. Weir, S. C., Fischer, S. H., Stock, F. & Gill, V. J. ( 1998;). Detection of Legionella by PCR in respiratory specimens using a commercially available kit. Am J Clin Pathol 110, 295–300.
    [Google Scholar]
  31. Winn, W. C., Jr, Davis, G. S., Gump, D. W., Craighead, J. E. & Beaty, H. N. ( 1982;). Legionnaires’ pneumonia after intratracheal inoculation of guinea pigs and rats. Lab Invest 47, 568–578.
    [Google Scholar]
  32. Yanagihara, K., Tomono, K., Sawai, T., Hirakata, Y., Kadota, J., Koga, H., Tashiro, T. & Kohno, S. ( 1997;). Effect of clarithromycin on lymphocytes in chronic respiratory Pseudomonas aeruginosa infection. Am J Respir Crit Care Med 155, 337–342.[CrossRef]
    [Google Scholar]
  33. Yu, V. L. (1995). Legionella pneumophila (Legionnaires’ disease). In Principles and Practice of Infectious Diseases, 4th edn, pp. 2087–2097. Edited by G. L. Mandell, J. E. Bennett & R. Doln. New York: Churchill Livingstone.
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