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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1992 Dec;30(12):3280–3283. doi: 10.1128/jcm.30.12.3280-3283.1992

Rapid, sensitive detection of Mycoplasma pneumoniae in simulated clinical specimens by DNA amplification.

G E Buck 1, L C O'Hara 1, J T Summersgill 1
PMCID: PMC270650  PMID: 1452717

Abstract

The polymerase chain reaction (PCR) was investigated as a means of diagnosing Mycoplasma pneumoniae infections. The target DNA sequence was a 375-bp segment of the P1 virulence protein. This DNA segment was amplified in pure cultures of five different strains of M. pneumoniae but not in other species of Mycoplasma, Acholeplasma, or Ureaplasma that were tested. Simulated clinical specimens were used to compare PCR, culture, and the gene probe. The sensitivity of PCR was between 1 and 10 organisms. The sensitivity of culture was approximately 10(3) organisms, and the gene probe detected between 10(4) and 10(5) organisms. These results indicate that PCR has significant potential as a rapid, sensitive method for detecting M. pneumoniae in clinical specimens.

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Selected References

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  1. Balassanian N., Robbins F. C. Mycoplasma pneumoniae infection in families. N Engl J Med. 1967 Oct 5;277(14):719–725. doi: 10.1056/NEJM196710052771401. [DOI] [PubMed] [Google Scholar]
  2. Bernet C., Garret M., de Barbeyrac B., Bebear C., Bonnet J. Detection of Mycoplasma pneumoniae by using the polymerase chain reaction. J Clin Microbiol. 1989 Nov;27(11):2492–2496. doi: 10.1128/jcm.27.11.2492-2496.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Buck G. E., O'Hara L. C., Summersgill J. T. Rapid, simple method for treating clinical specimens containing Mycobacterium tuberculosis to remove DNA for polymerase chain reaction. J Clin Microbiol. 1992 May;30(5):1331–1334. doi: 10.1128/jcm.30.5.1331-1334.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Clyde W. A., Jr Mycoplasma pneumoniae respiratory disease symposium: summation and significance. Yale J Biol Med. 1983 Sep-Dec;56(5-6):523–527. [PMC free article] [PubMed] [Google Scholar]
  5. Davis C. P., Cochran S., Lisse J., Buck G., DiNuzzo A. R., Weber T., Reinarz J. A. Isolation of Mycoplasma pneumoniae from synovial fluid samples in a patient with pneumonia and polyarthritis. Arch Intern Med. 1988 Apr;148(4):969–970. [PubMed] [Google Scholar]
  6. Dular R., Kajioka R., Kasatiya S. Comparison of Gen-Probe commercial kit and culture technique for the diagnosis of Mycoplasma pneumoniae infection. J Clin Microbiol. 1988 May;26(5):1068–1069. doi: 10.1128/jcm.26.5.1068-1069.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dussaix E., Slim A., Tournier P. Comparison of enzyme-linked immunosorbent assay (ELISA) and complement fixation test for detection of Mycoplasma pneumoniae antibodies. J Clin Pathol. 1983 Feb;36(2):228–232. doi: 10.1136/jcp.36.2.228. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. ENNY G. E., GRAYSTON J. T. EATON PLEUROPNEUMONIA-LIKE ORGANISM (MYCOPLASMA PNEUMONIAE) COMPLEMENT-FIXING ANTIGEN: EXTRACTION WITH ORGANIC SOLVENTS. J Immunol. 1965 Jul;95:19–25. [PubMed] [Google Scholar]
  9. Fernald G. W., Collier A. M., Clyde W. A., Jr Respiratory infections due to Mycoplasma pneumoniae in infants and children. Pediatrics. 1975 Mar;55(3):327–335. [PubMed] [Google Scholar]
  10. Foy H. M., Allan I. D. Frequency of Mycoplasma pneumoniae pneumonia. Lancet. 1982 Feb 13;1(8268):392–392. doi: 10.1016/s0140-6736(82)91413-1. [DOI] [PubMed] [Google Scholar]
  11. Foy H. M., Cooney M. K., McMahan R., Grayston J. T. Viral and mycoplasmal pneumonia in a prepaid medical care group during an eight-year period. Am J Epidemiol. 1973 Feb;97(2):93–102. doi: 10.1093/oxfordjournals.aje.a121493. [DOI] [PubMed] [Google Scholar]
  12. Foy H. M., Kenny G. E., Cooney M. K., Allan I. D. Long-term epidemiology of infections with Mycoplasma pneumoniae. J Infect Dis. 1979 Jun;139(6):681–687. doi: 10.1093/infdis/139.6.681. [DOI] [PubMed] [Google Scholar]
  13. Harris R., Marmion B. P., Varkanis G., Kok T., Lunn B., Martin J. Laboratory diagnosis of Mycoplasma pneumoniae infection. 2. Comparison of methods for the direct detection of specific antigen or nucleic acid sequences in respiratory exudates. Epidemiol Infect. 1988 Dec;101(3):685–694. doi: 10.1017/s0950268800029563. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hata D., Kuze F., Mochizuki Y., Ohkubo H., Kanazashi S., Maeda S., Miwa N., Mikawa H. Evaluation of DNA probe test for rapid diagnosis of Mycoplasma pneumoniae infections. J Pediatr. 1990 Feb;116(2):273–276. doi: 10.1016/s0022-3476(05)82889-6. [DOI] [PubMed] [Google Scholar]
  15. Jensen J. S., Søndergård-Andersen J., Uldum S. A., Lind K. Detection of Mycoplasma pneumoniae in simulated clinical samples by polymerase chain reaction. Brief report. APMIS. 1989 Nov;97(11):1046–1048. doi: 10.1111/j.1699-0463.1989.tb00516.x. [DOI] [PubMed] [Google Scholar]
  16. Kingery F. A. When an ear nodule hurts. JAMA. 1966 Jul 11;197(2):137–137. [PubMed] [Google Scholar]
  17. Kleemola S. R., Karjalainen J. E., Räty R. K. Rapid diagnosis of Mycoplasma pneumoniae infection: clinical evaluation of a commercial probe test. J Infect Dis. 1990 Jul;162(1):70–75. doi: 10.1093/infdis/162.1.70. [DOI] [PubMed] [Google Scholar]
  18. Mizutani H., Mizutani H. Immunologic responses in patients with Mycoplasma pneumoniae infections. Am Rev Respir Dis. 1983 Feb;127(2):175–179. doi: 10.1164/arrd.1983.127.2.175. [DOI] [PubMed] [Google Scholar]
  19. Murray H. W., Masur H., Senterfit L. B., Roberts R. B. The protean manifestations of Mycoplasma pneumoniae infection in adults. Am J Med. 1975 Feb;58(2):229–242. doi: 10.1016/0002-9343(75)90574-4. [DOI] [PubMed] [Google Scholar]
  20. Persing D. H. Polymerase chain reaction: trenches to benches. J Clin Microbiol. 1991 Jul;29(7):1281–1285. doi: 10.1128/jcm.29.7.1281-1285.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  22. Siegler D. I. Lung abscess associated with Mycoplasma pneumoniae infection. Br J Dis Chest. 1973 Apr;67(2):123–127. doi: 10.1016/0007-0971(73)90041-7. [DOI] [PubMed] [Google Scholar]
  23. Sillis M. The limitations of IgM assays in the serological diagnosis of Mycoplasma pneumoniae infections. J Med Microbiol. 1990 Dec;33(4):253–258. doi: 10.1099/00222615-33-4-253. [DOI] [PubMed] [Google Scholar]
  24. Su C. J., Tryon V. V., Baseman J. B. Cloning and sequence analysis of cytadhesin P1 gene from Mycoplasma pneumoniae. Infect Immun. 1987 Dec;55(12):3023–3029. doi: 10.1128/iai.55.12.3023-3029.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Tilton R. C., Dias F., Kidd H., Ryan R. W. DNA probe versus culture for detection of Mycoplasma pneumoniae in clinical specimens. Diagn Microbiol Infect Dis. 1988 Jun;10(2):109–112. doi: 10.1016/0732-8893(88)90048-x. [DOI] [PubMed] [Google Scholar]
  26. Tully J. G., Rose D. L., Whitcomb R. F., Wenzel R. P. Enhanced isolation of Mycoplasma pneumoniae from throat washings with a newly-modified culture medium. J Infect Dis. 1979 Apr;139(4):478–482. doi: 10.1093/infdis/139.4.478. [DOI] [PubMed] [Google Scholar]
  27. Uldum S. A., Jensen J. S., Søndergård-Andersen J., Lind K. Enzyme immunoassay for detection of immunoglobulin M (IgM) and IgG antibodies to Mycoplasma pneumoniae. J Clin Microbiol. 1992 May;30(5):1198–1204. doi: 10.1128/jcm.30.5.1198-1204.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. White R. J., Blainey A. D., Harrison K. J., Clarke S. K. Causes of pneumonia presenting to a district general hospital. Thorax. 1981 Aug;36(8):566–570. doi: 10.1136/thx.36.8.566. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. van Griethuysen A. J., de Graaf R., van Druten J. A., Heessen F. W., van der Logt J. T., van Loon A. M. Use of the enzyme-linked immunosorbent assay for the early diagnosis of Mycoplasma pneumoniae infection. Eur J Clin Microbiol. 1984 Apr;3(2):116–121. doi: 10.1007/BF02014328. [DOI] [PubMed] [Google Scholar]

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