Abstract
A new molecular assay, based on a rapid DNA extraction protocol, PCR, and hybridization to a specific probe in a nonradioactive microwell plate format was used to detect Mycoplasma pneumoniae in bronchoalveolar fluid specimens. The sensitivity of the assay was determined to be 10 to 100 organisms with M. pneumoniae reference strains. Specificity testing with different bacteria capable of producing pneumonia showed no cross-reactivity. In a prospective study, bronchoalveolar lavage fluids obtained from patients with pneumonia were investigated with the PCR assay and compared to culture. Twelve positive samples were detected with the PCR assay. Seven of them were subsequently confirmed by culture. All patients with positive PCR results seroconverted. Application of the PCR assay described may lead to safe and early diagnosis of M. pneumoniae in patients with pneumonia.
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- 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]
- Blackmore T. K., Reznikov M., Gordon D. L. Clinical utility of the polymerase chain reaction to diagnose Mycoplasma pneumoniae infection. Pathology. 1995 Apr;27(2):177–181. doi: 10.1080/00313029500169832. [DOI] [PubMed] [Google Scholar]
- Buck G. E., O'Hara L. C., Summersgill J. T. Rapid, sensitive detection of Mycoplasma pneumoniae in simulated clinical specimens by DNA amplification. J Clin Microbiol. 1992 Dec;30(12):3280–3283. doi: 10.1128/jcm.30.12.3280-3283.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cassol S., Butcher A., Kinard S., Spadoro J., Sy T., Lapointe N., Read S., Gomez P., Fauvel M., Major C. Rapid screening for early detection of mother-to-child transmission of human immunodeficiency virus type 1. J Clin Microbiol. 1994 Nov;32(11):2641–2645. doi: 10.1128/jcm.32.11.2641-2645.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- Kessler H. H., Pierer K., Stuenzner D., Auer-Grumbach P., Haller E. M., Marth E. Rapid detection of Chlamydia trachomatis in conjunctival, pharyngeal, and urethral specimens with a new polymerase chain reaction assay. Sex Transm Dis. 1994 Jul-Aug;21(4):191–195. doi: 10.1097/00007435-199407000-00002. [DOI] [PubMed] [Google Scholar]
- Kessler H. H., Reinthaler F. F., Pschaid A., Pierer K., Kleinhappl B., Eber E., Marth E. Rapid detection of Legionella species in bronchoalveolar lavage fluids with the EnviroAmp Legionella PCR amplification and detection kit. J Clin Microbiol. 1993 Dec;31(12):3325–3328. doi: 10.1128/jcm.31.12.3325-3328.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Loeffelholz M. J., Lewinski C. A., Silver S. R., Purohit A. P., Herman S. A., Buonagurio D. A., Dragon E. A. Detection of Chlamydia trachomatis in endocervical specimens by polymerase chain reaction. J Clin Microbiol. 1992 Nov;30(11):2847–2851. doi: 10.1128/jcm.30.11.2847-2851.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lüneberg E., Jensen J. S., Frosch M. Detection of Mycoplasma pneumoniae by polymerase chain reaction and nonradioactive hybridization in microtiter plates. J Clin Microbiol. 1993 May;31(5):1088–1094. doi: 10.1128/jcm.31.5.1088-1094.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Marmion B. P., Williamson J., Worswick D. A., Kok T. W., Harris R. J. Experience with newer techniques for the laboratory detection of Mycoplasma pneumoniae infection: Adelaide, 1978-1992. Clin Infect Dis. 1993 Aug;17 (Suppl 1):S90–S99. doi: 10.1093/clinids/17.supplement_1.s90. [DOI] [PubMed] [Google Scholar]
- Nolte F. S., Thurmond C., Fried M. W. Preclinical evaluation of AMPLICOR hepatitis C virus test for detection of hepatitis C virus RNA. J Clin Microbiol. 1995 Jul;33(7):1775–1778. doi: 10.1128/jcm.33.7.1775-1778.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Skakni L., Sardet A., Just J., Landman-Parker J., Costil J., Moniot-Ville N., Bricout F., Garbarg-Chenon A. Detection of Mycoplasma pneumoniae in clinical samples from pediatric patients by polymerase chain reaction. J Clin Microbiol. 1992 Oct;30(10):2638–2643. doi: 10.1128/jcm.30.10.2638-2643.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tjhie J. H., van Kuppeveld F. J., Roosendaal R., Melchers W. J., Gordijn R., MacLaren D. M., Walboomers J. M., Meijer C. J., van den Brule A. J. Direct PCR enables detection of Mycoplasma pneumoniae in patients with respiratory tract infections. J Clin Microbiol. 1994 Jan;32(1):11–16. doi: 10.1128/jcm.32.1.11-16.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vekris A., Bauduer F., Maillet S., Bébéar C., Bonnet J. Improved microplate immunoenzymatic assay of PCR products for rapid detection of Mycoplasma pneumoniae. Mol Cell Probes. 1995 Feb;9(1):25–31. doi: 10.1016/s0890-8508(95)90947-8. [DOI] [PubMed] [Google Scholar]
- Vignoli C., de Lamballerie X., Zandotti C., Tamalet C., de Micco P. Advantage of a rapid extraction method of HIV1 DNA suitable for polymerase chain reaction. Res Virol. 1995 Mar-Apr;146(2):159–162. doi: 10.1016/0923-2516(96)81085-5. [DOI] [PubMed] [Google Scholar]
- Zandotti C., De Lamballerie X., Guignole-Vignoli C., Bollet C., De Micco P. A rapid DNA extraction method from culture and clinical samples. Suitable for the detection of human cytomegalovirus by the polymerase chain reaction. Acta Virol. 1993 Feb;37(1):106–108. [PubMed] [Google Scholar]
- de Lamballerie X., Zandotti C., Vignoli C., Bollet C., de Micco P. A one-step microbial DNA extraction method using "Chelex 100" suitable for gene amplification. Res Microbiol. 1992 Oct;143(8):785–790. doi: 10.1016/0923-2508(92)90107-y. [DOI] [PubMed] [Google Scholar]