Skip to main content
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1997 Oct;35(10):2503–2510. doi: 10.1128/jcm.35.10.2503-2510.1997

Use of different molecular typing techniques for bacteriological follow-up in a clinical trial with AIDS patients with Mycobacterium avium bacteremia.

M Picardeau 1, A Varnerot 1, T Lecompte 1, F Brel 1, T May 1, V Vincent 1
PMCID: PMC230000  PMID: 9316897

Abstract

One hundred ninety-six Mycobacterium avium isolates from blood samples recovered from 93 AIDS patients for several months were typed by serotyping, by IS1245 restriction fragment length polymorphism (RFLP) analysis and in some cases RFLP analysis with plasmids pVT2 and pLR7 as probes, and by pulsed-field gel electrophoresis (PFGE). PCR typing of single colonies was also used to detect polyclonal infections. Strains belonged mainly to serotypes 1, 4, and 8. pVT2- and pLR7-related plasmids were detected in strains from 49% of the patients. The IS1245 RFLP and PFGE analyses showed a 96.8% diversity of the M. avium strains from the 93 patients. The vast majority (95.2%) of infections were monoclonal, indicating that recent infection is unlikely, even at an advanced stage of AIDS. For one patient, sequential isolates gave divergent patterns of sensitivity and resistance to clarithromycin, but all were identified as the initial clone. RFLP analysis and PCR typing of single colonies allowed for the detection of three polyclonal infections during the bacteriological follow-up. Among strains from patients whose samples were positive by culture after treatment for 2 to 15 months, 97.4% were the same as the initial strain. In conclusion, relapses and failures were mostly due to the initial strain. These relapses and failures resulted either from the selection of resistant mutants or the reappearance of sensitive strains, suggesting the persistence of nonsterilized tissue reservoirs.

Full Text

The Full Text of this article is available as a PDF (1.6 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arbeit R. D., Slutsky A., Barber T. W., Maslow J. N., Niemczyk S., Falkinham J. O., 3rd, O'Connor G. T., von Reyn C. F. Genetic diversity among strains of Mycobacterium avium causing monoclonal and polyclonal bacteremia in patients with AIDS. J Infect Dis. 1993 Jun;167(6):1384–1390. doi: 10.1093/infdis/167.6.1384. [DOI] [PubMed] [Google Scholar]
  2. Askgaard D. S., Giese S. B., Thybo S., Lerche A., Bennedsen J. Serovars of Mycobacterium avium complex isolated from patients in Denmark. J Clin Microbiol. 1994 Nov;32(11):2880–2882. doi: 10.1128/jcm.32.11.2880-2882.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brennan P. J., Heifets M., Ullom B. P. Thin-layer chromatography of lipid antigens as a means of identifying nontuberculous mycobacteria. J Clin Microbiol. 1982 Mar;15(3):447–455. doi: 10.1128/jcm.15.3.447-455.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cave M. D., Eisenach K. D., Templeton G., Salfinger M., Mazurek G., Bates J. H., Crawford J. T. Stability of DNA fingerprint pattern produced with IS6110 in strains of Mycobacterium tuberculosis. J Clin Microbiol. 1994 Jan;32(1):262–266. doi: 10.1128/jcm.32.1.262-266.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Crawford J. T., Bates J. H. Analysis of plasmids in Mycobacterium avium-intracellulare isolates from persons with acquired immunodeficiency syndrome. Am Rev Respir Dis. 1986 Oct;134(4):659–661. doi: 10.1164/arrd.1986.134.4.659. [DOI] [PubMed] [Google Scholar]
  6. Dawson D. J. Infection with Mycobacterium avium complex in Australian patients with AIDS. Med J Aust. 1990 Oct 15;153(8):466–468. doi: 10.5694/j.1326-5377.1990.tb126152.x. [DOI] [PubMed] [Google Scholar]
  7. De Smet K. A., Brown I. N., Yates M., Ivanyi J. Ribosomal internal transcribed spacer sequences are identical among Mycobacterium avium-intracellulare complex isolates from AIDS patients, but vary among isolates from elderly pulmonary disease patients. Microbiology. 1995 Oct;141(Pt 10):2739–2747. doi: 10.1099/13500872-141-10-2739. [DOI] [PubMed] [Google Scholar]
  8. De Smet K. A., Hellyer T. J., Khan A. W., Brown I. N., Ivanyi J. Genetic and serovar typing of clinical isolates of the Mycobacterium avium-intracellulare complex. Tuber Lung Dis. 1996 Feb;77(1):71–76. doi: 10.1016/s0962-8479(96)90079-9. [DOI] [PubMed] [Google Scholar]
  9. Falkinham J. O., 3rd Epidemiology of infection by nontuberculous mycobacteria. Clin Microbiol Rev. 1996 Apr;9(2):177–215. doi: 10.1128/cmr.9.2.177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Frothingham R., Wilson K. H. Molecular phylogeny of the Mycobacterium avium complex demonstrates clinically meaningful divisions. J Infect Dis. 1994 Feb;169(2):305–312. doi: 10.1093/infdis/169.2.305. [DOI] [PubMed] [Google Scholar]
  11. Guerrero C., Bernasconi C., Burki D., Bodmer T., Telenti A. A novel insertion element from Mycobacterium avium, IS1245, is a specific target for analysis of strain relatedness. J Clin Microbiol. 1995 Feb;33(2):304–307. doi: 10.1128/jcm.33.2.304-307.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hampson S. J., Portaels F., Thompson J., Green E. P., Moss M. T., Hermon-Taylor J., McFadden J. J. DNA probes demonstrate a single highly conserved strain of Mycobacterium avium infecting AIDS patients. Lancet. 1989 Jan 14;1(8629):65–68. doi: 10.1016/s0140-6736(89)91427-x. [DOI] [PubMed] [Google Scholar]
  13. Hellyer T. J., Brown I. N., Dale J. W., Easmon C. S. Plasmid analysis of Mycobacterium avium-intracellulare (MAI) isolated in the United Kingdom from patients with and without AIDS. J Med Microbiol. 1991 Apr;34(4):225–231. doi: 10.1099/00222615-34-4-225. [DOI] [PubMed] [Google Scholar]
  14. Horsburgh C. R., Jr, Cohn D. L., Roberts R. B., Masur H., Miller R. A., Tsang A. Y., Iseman M. D. Mycobacterium avium-M. intracellulare isolates from patients with or without acquired immunodeficiency syndrome. Antimicrob Agents Chemother. 1986 Dec;30(6):955–957. doi: 10.1128/aac.30.6.955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Horsburgh C. R., Jr Mycobacterium avium complex infection in the acquired immunodeficiency syndrome. N Engl J Med. 1991 May 9;324(19):1332–1338. doi: 10.1056/NEJM199105093241906. [DOI] [PubMed] [Google Scholar]
  16. Inderlied C. B., Kemper C. A., Bermudez L. E. The Mycobacterium avium complex. Clin Microbiol Rev. 1993 Jul;6(3):266–310. doi: 10.1128/cmr.6.3.266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jucker M. T., Falkinham J. O., 3rd Epidemiology of infection by nontuberculous mycobacteria IX. Evidence for two DNA homology groups among small plasmids in Mycobacterium avium, Mycobacterium intracellulare, and Mycobacterium scrofulaceum. Am Rev Respir Dis. 1990 Oct;142(4):858–862. doi: 10.1164/ajrccm/142.4.858. [DOI] [PubMed] [Google Scholar]
  18. Julander I., Hoffner S., Petrini B., Ostlund L. Multiple serovars of Mycobacterium avium complex in patients with AIDS. APMIS. 1996 Apr;104(4):318–320. [PubMed] [Google Scholar]
  19. Kemper C. A., Havlir D., Bartok A. E., Kane C., Camp B., Lane N., Deresinski S. C. Transient bacteremia due to Mycobacterium avium complex in patients with AIDS. J Infect Dis. 1994 Aug;170(2):488–493. doi: 10.1093/infdis/170.2.488. [DOI] [PubMed] [Google Scholar]
  20. Meier A., Heifets L., Wallace R. J., Jr, Zhang Y., Brown B. A., Sander P., Böttger E. C. Molecular mechanisms of clarithromycin resistance in Mycobacterium avium: observation of multiple 23S rDNA mutations in a clonal population. J Infect Dis. 1996 Aug;174(2):354–360. doi: 10.1093/infdis/174.2.354. [DOI] [PubMed] [Google Scholar]
  21. Meier A., Kirschner P., Springer B., Steingrube V. A., Brown B. A., Wallace R. J., Jr, Böttger E. C. Identification of mutations in 23S rRNA gene of clarithromycin-resistant Mycobacterium intracellulare. Antimicrob Agents Chemother. 1994 Feb;38(2):381–384. doi: 10.1128/aac.38.2.381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Morris S. L., Rouse D. A., Malik A., Chaparas S. D., Witebsky F. G. Characterisation of plasmids extracted from AIDS--associated Mycobacterium avium isolates. Tubercle. 1990 Sep;71(3):181–185. doi: 10.1016/0041-3879(90)90073-h. [DOI] [PubMed] [Google Scholar]
  23. Nash K. A., Inderlied C. B. Genetic basis of macrolide resistance in Mycobacterium avium isolated from patients with disseminated disease. Antimicrob Agents Chemother. 1995 Dec;39(12):2625–2630. doi: 10.1128/aac.39.12.2625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Picardeau M., Varnerot A., Rauzier J., Gicquel B., Vincent V. Mycobacterium xenopi IS1395, a novel insertion sequence expanding the IS256 family. Microbiology. 1996 Sep;142(Pt 9):2453–2461. doi: 10.1099/00221287-142-9-2453. [DOI] [PubMed] [Google Scholar]
  25. Picardeau M., Vincent V. Typing of Mycobacterium avium isolates by PCR. J Clin Microbiol. 1996 Feb;34(2):389–392. doi: 10.1128/jcm.34.2.389-392.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Portaels F. Epidemiology of mycobacterial diseases. Clin Dermatol. 1995 May-Jun;13(3):207–222. doi: 10.1016/0738-081x(95)00004-y. [DOI] [PubMed] [Google Scholar]
  27. Prinzis S., Rivoire B., Brennan P. J. Search for the molecular basis of morphological variation in Mycobacterium avium. Infect Immun. 1994 May;62(5):1946–1951. doi: 10.1128/iai.62.5.1946-1951.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Roiz M. P., Palenque E., Guerrero C., Garcia M. J. Use of restriction fragment length polymorphism as a genetic marker for typing Mycobacterium avium strains. J Clin Microbiol. 1995 May;33(5):1389–1391. doi: 10.1128/jcm.33.5.1389-1391.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Slutsky A. M., Arbeit R. D., Barber T. W., Rich J., von Reyn C. F., Pieciak W., Barlow M. A., Maslow J. N. Polyclonal infections due to Mycobacterium avium complex in patients with AIDS detected by pulsed-field gel electrophoresis of sequential clinical isolates. J Clin Microbiol. 1994 Jul;32(7):1773–1778. doi: 10.1128/jcm.32.7.1773-1778.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tenover F. C., Arbeit R. D., Goering R. V., Mickelsen P. A., Murray B. E., Persing D. H., Swaminathan B. Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing. J Clin Microbiol. 1995 Sep;33(9):2233–2239. doi: 10.1128/jcm.33.9.2233-2239.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Thierry D., Vincent V., Clément F., Guesdon J. L. Isolation of specific DNA fragments of Mycobacterium avium and their possible use in diagnosis. J Clin Microbiol. 1993 May;31(5):1048–1054. doi: 10.1128/jcm.31.5.1048-1054.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Torriani F. J., Behling C. A., McCutchan J. A., Haubrich R. H., Havlir D. V. Disseminated Mycobacterium avium complex: correlation between blood and tissue burden. J Infect Dis. 1996 Apr;173(4):942–949. doi: 10.1093/infdis/173.4.942. [DOI] [PubMed] [Google Scholar]
  33. Tsang A. Y., Denner J. C., Brennan P. J., McClatchy J. K. Clinical and epidemiological importance of typing of Mycobacterium avium complex isolates. J Clin Microbiol. 1992 Feb;30(2):479–484. doi: 10.1128/jcm.30.2.479-484.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Varadaraj K., Skinner D. M. Denaturants or cosolvents improve the specificity of PCR amplification of a G + C-rich DNA using genetically engineered DNA polymerases. Gene. 1994 Mar 11;140(1):1–5. doi: 10.1016/0378-1119(94)90723-4. [DOI] [PubMed] [Google Scholar]
  35. Wayne L. G., Young L. S., Bertram M. Absence of mycobacterial antibody in patients with acquired immune deficiency syndrome. Eur J Clin Microbiol. 1986 Jun;5(3):363–365. doi: 10.1007/BF02017802. [DOI] [PubMed] [Google Scholar]
  36. Wright E. L., Zywno-van Ginkel S., Rastogi N., Barrow W. W. Monoclonal infection involving Mycobacterium avium presenting with three distinct colony morphotypes. J Clin Microbiol. 1996 Oct;34(10):2475–2478. doi: 10.1128/jcm.34.10.2475-2478.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Yajko D. M., Chin D. P., Gonzalez P. C., Nassos P. S., Hopewell P. C., Reingold A. L., Horsburgh C. R., Jr, Yakrus M. A., Ostroff S. M., Hadley W. K. Mycobacterium avium complex in water, food, and soil samples collected from the environment of HIV-infected individuals. J Acquir Immune Defic Syndr Hum Retrovirol. 1995 Jun 1;9(2):176–182. [PubMed] [Google Scholar]
  38. Yakrus M. A., Good R. C. Geographic distribution, frequency, and specimen source of Mycobacterium avium complex serotypes isolated from patients with acquired immunodeficiency syndrome. J Clin Microbiol. 1990 May;28(5):926–929. doi: 10.1128/jcm.28.5.926-929.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. von Reyn C. F., Barber T. W., Arbeit R. D., Sox C. H., O'Connor G. T., Brindle R. J., Gilks C. F., Hakkarainen K., Ranki A., Bartholomew C. Evidence of previous infection with Mycobacterium avium-Mycobacterium intracellulare complex among healthy subjects: an international study of dominant mycobacterial skin test reactions. J Infect Dis. 1993 Dec;168(6):1553–1558. doi: 10.1093/infdis/168.6.1553. [DOI] [PubMed] [Google Scholar]
  40. von Reyn C. F., Jacobs N. J., Arbeit R. D., Maslow J. N., Niemczyk S. Polyclonal Mycobacterium avium infections in patients with AIDS: variations in antimicrobial susceptibilities of different strains of M. avium isolated from the same patient. J Clin Microbiol. 1995 Apr;33(4):1008–1010. doi: 10.1128/jcm.33.4.1008-1010.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. von Reyn C. F., Maslow J. N., Barber T. W., Falkinham J. O., 3rd, Arbeit R. D. Persistent colonisation of potable water as a source of Mycobacterium avium infection in AIDS. Lancet. 1994 May 7;343(8906):1137–1141. doi: 10.1016/s0140-6736(94)90239-9. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES