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. 1993 Jul;31(7):1677–1682. doi: 10.1128/jcm.31.7.1677-1682.1993

Molecular strain typing of Mycobacterium tuberculosis to confirm cross-contamination in the mycobacteriology laboratory and modification of procedures to minimize occurrence of false-positive cultures.

P M Small 1, N B McClenny 1, S P Singh 1, G K Schoolnik 1, L S Tompkins 1, P A Mickelsen 1
PMCID: PMC265613  PMID: 8102372

Abstract

Molecular strain typing by restriction fragment length polymorphism analysis was used to demonstrate that two clusters of Mycobacterium tuberculosis cultures involving six patients resulted from cross-contamination in the mycobacteriology laboratory. Contaminated cultures were processed by the decontamination procedure and were read on the BACTEC instrument following acid-fast bacillus smear-positive specimens from patients with active tuberculosis. Investigation of these episodes suggested opportunities for modification of laboratory procedures to minimize cross-contamination and confirmed the adverse medical and public health consequences of false-positive cultures. Strain-typing results were used in decisions regarding patient care, including the curtailment of unnecessary treatment in one patient. Molecular strain typing appears to be a valuable means of identifying false-positive cultures of M. tuberculosis in selected settings.

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Selected References

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  1. Aber V. R., Allen B. W., Mitchison D. A., Ayuma P., Edwards E. A., Keyes A. B. Quality control in tuberculosis bacteriology. 1. Laboratory studies on isolated positive cultures and the efficiency of direct smear examination. Tubercle. 1980 Sep;61(3):123–133. doi: 10.1016/0041-3879(80)90001-x. [DOI] [PubMed] [Google Scholar]
  2. Barnes P. F., Bloch A. B., Davidson P. T., Snider D. E., Jr Tuberculosis in patients with human immunodeficiency virus infection. N Engl J Med. 1991 Jun 6;324(23):1644–1650. doi: 10.1056/NEJM199106063242307. [DOI] [PubMed] [Google Scholar]
  3. Conville P. S., Witebsky F. G. Inter-bottle transfer of mycobacteria by the BACTEC 460. Diagn Microbiol Infect Dis. 1989 Sep-Oct;12(5):401–405. doi: 10.1016/0732-8893(89)90110-7. [DOI] [PubMed] [Google Scholar]
  4. Dizon D., Mihailescu C., Bae H. C. Simple procedure for detection of Mycobacterium gordonae in water causing false-positive acid-fast smears. J Clin Microbiol. 1976 Feb;3(2):211–211. doi: 10.1128/jcm.3.2.211-211.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dooley S. W., Jarvis W. R., Martone W. J., Snider D. E., Jr Multidrug-resistant tuberculosis. Ann Intern Med. 1992 Aug 1;117(3):257–259. doi: 10.7326/0003-4819-117-3-257. [DOI] [PubMed] [Google Scholar]
  6. Fischl M. A., Uttamchandani R. B., Daikos G. L., Poblete R. B., Moreno J. N., Reyes R. R., Boota A. M., Thompson L. M., Cleary T. J., Lai S. An outbreak of tuberculosis caused by multiple-drug-resistant tubercle bacilli among patients with HIV infection. Ann Intern Med. 1992 Aug 1;117(3):177–183. doi: 10.7326/0003-4819-117-3-177. [DOI] [PubMed] [Google Scholar]
  7. Graham L., Jr, Warren N. G., Tsang A. Y., Dalton H. P. Mycobacterium avium complex pseudobacteriuria from a hospital water supply. J Clin Microbiol. 1988 May;26(5):1034–1036. doi: 10.1128/jcm.26.5.1034-1036.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gubler J. G., Salfinger M., von Graevenitz A. Pseudoepidemic of nontuberculous mycobacteria due to a contaminated bronchoscope cleaning machine. Report of an outbreak and review of the literature. Chest. 1992 May;101(5):1245–1249. doi: 10.1378/chest.101.5.1245. [DOI] [PubMed] [Google Scholar]
  9. Hermans P. W., van Soolingen D., Dale J. W., Schuitema A. R., McAdam R. A., Catty D., van Embden J. D. Insertion element IS986 from Mycobacterium tuberculosis: a useful tool for diagnosis and epidemiology of tuberculosis. J Clin Microbiol. 1990 Sep;28(9):2051–2058. doi: 10.1128/jcm.28.9.2051-2058.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jones W. D., Jr Bacteriophage typing of Mycobacterium tuberculosis cultures from incidents of suspected laboratory cross-contamination. Tubercle. 1988 Mar;69(1):43–46. doi: 10.1016/0041-3879(88)90039-6. [DOI] [PubMed] [Google Scholar]
  11. MacGregor R. R., Clark L. W., Bass F. The significance of isolating low numbers of mycobacterium tuberculosis in culture of sputum specimens. Chest. 1975 Oct;68(4):518–523. doi: 10.1378/chest.68.4.518. [DOI] [PubMed] [Google Scholar]
  12. Maurer J. R., Desmond E. P., Lesser M. D., Jones W. D., Jr False-positive cultures of Mycobacterium tuberculosis. Chest. 1984 Sep;86(3):439–443. doi: 10.1378/chest.86.3.439. [DOI] [PubMed] [Google Scholar]
  13. Mitchison D. A., Keyes A. B., Edwards E. A., Ayuma P., Byfield S. P., Nunn A. J. Quality control in tuberculosis bacteriology. 2. The origin of isolated positive cultures from the sputum of patients in four studies of short course chemotherapy in Africa. Tubercle. 1980 Sep;61(3):135–144. doi: 10.1016/0041-3879(80)90002-1. [DOI] [PubMed] [Google Scholar]
  14. Murray P. R. Mycobacterial cross-contamination with the modified BACTEC 460 TB System. Diagn Microbiol Infect Dis. 1991 Jan-Feb;14(1):33–35. doi: 10.1016/0732-8893(91)90085-t. [DOI] [PubMed] [Google Scholar]
  15. Small P. M., Shafer R. W., Hopewell P. C., Singh S. P., Murphy M. J., Desmond E., Sierra M. F., Schoolnik G. K. Exogenous reinfection with multidrug-resistant Mycobacterium tuberculosis in patients with advanced HIV infection. N Engl J Med. 1993 Apr 22;328(16):1137–1144. doi: 10.1056/NEJM199304223281601. [DOI] [PubMed] [Google Scholar]
  16. Smith W. B., Vance D. W., Jr Specimen cross-contamination by a strain of Mycobacterium tuberculosis lacking nitrate reductase activity. Diagn Microbiol Infect Dis. 1991 Nov-Dec;14(6):523–526. doi: 10.1016/0732-8893(91)90012-5. [DOI] [PubMed] [Google Scholar]
  17. Snider D. E., Jr, Roper W. L. The new tuberculosis. N Engl J Med. 1992 Mar 5;326(10):703–705. doi: 10.1056/NEJM199203053261011. [DOI] [PubMed] [Google Scholar]
  18. Vannier A. M., Tarrand J. J., Murray P. R. Mycobacterial cross contamination during radiometric culturing. J Clin Microbiol. 1988 Sep;26(9):1867–1868. doi: 10.1128/jcm.26.9.1867-1868.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Witebsky F. G., Keiser J. F., Conville P. S., Bryan R., Park C. H., Walker R., Siddiqi S. H. Comparison of BACTEC 13A medium and Du Pont isolator for detection of mycobacteremia. J Clin Microbiol. 1988 Aug;26(8):1501–1505. doi: 10.1128/jcm.26.8.1501-1505.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. van Embden J. D., Cave M. D., Crawford J. T., Dale J. W., Eisenach K. D., Gicquel B., Hermans P., Martin C., McAdam R., Shinnick T. M. Strain identification of Mycobacterium tuberculosis by DNA fingerprinting: recommendations for a standardized methodology. J Clin Microbiol. 1993 Feb;31(2):406–409. doi: 10.1128/jcm.31.2.406-409.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. van Embden J. D., van Soolingen D., Small P. M., Hermans P. W. Genetic markers for the epidemiology of tuberculosis. Res Microbiol. 1992 May;143(4):385–391. doi: 10.1016/0923-2508(92)90051-o. [DOI] [PubMed] [Google Scholar]

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