1887

Abstract

Summary

A DNA amplification method was used to detect in clinical samples. 16S ribosomal RNA gene sequences were selected as the amplification target region. The polymerase chain reaction (PCR) with purified DNA fragments as templates yielded an expected 88-bp fragment from but not from other spp. nor from any of the other bacteria assayed. With this method, the 88-bp product specific for could be obtained from a minimum of 0.05 pg of DNA. Subsequently this PCR technique was used for the detection of in throat-swab samples. Twenty-two of 30 culture-positive clinical samples gave positive results in the PCR test. Thirty-two culture-negative clinical samples and 33 samples from healthy volunteers, of which only one was culture-positive, gave negative results in the same PCR test. This PCR method is useful for the direct detection of in clinical samples.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/00222615-38-3-166
1993-03-01
2024-07-22
Loading full text...

Full text loading...

/deliver/fulltext/jmm/38/3/medmicro-38-3-166.html?itemId=/content/journal/jmm/10.1099/00222615-38-3-166&mimeType=html&fmt=ahah

References

  1. Eaton MD, Meiklejohn G, van Herick W. Studies on the etiology of primary atypical pneumonia; a filtrable agent transmissible to cotton rats, hamsters and chick embryos. J Exp Med 1944; 79:649–668
    [Google Scholar]
  2. Denny FW, Clyde WA, Glezen WP. Mycoplasma pneumoniae disease: clinical spectrum, pathophysiology, epidemiology, and control. J Infect Dis 1971; 123:74–92
    [Google Scholar]
  3. Kenny GE, Grayston JT. Eaton pleuropneumoniae-like or ganism (Mycoplasma pneumoniae) complement-fixing antigen: extraction with organic solvents. J Immunol 1965; 95:19–25
    [Google Scholar]
  4. Dowdle WR, Robinson RQ. An indirect hemagglutination test for diagnosis of Mycoplasma pneumoniae infections. Proc Soc Exp Biol Med 1964; 116:947–950
    [Google Scholar]
  5. Jacobs E, Bennewitz A, Bredt W. Reaction pattern of human anti-Mycoplasma pneumoniae antibodies in enzyme-linked immunosorbent assays and immunoblotting. J Clin Microbiol 1986; 23:517–522
    [Google Scholar]
  6. Göbel UB, Geiser A, Stanbridge EJ. Oligonucleotide probes complementary to variable regions of ribosomal RNA discriminate between Mycoplasma species. J Gen Microbiol 1987; 133:1969–1974
    [Google Scholar]
  7. Hyman HC, Yogev D, Razin S. DNA probes for detection and identification of Mycoplasma pneumoniae and Mycoplasma genitalium. J Clin Microbiol 1987; 25:726–728
    [Google Scholar]
  8. 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; 26:1068–1069
    [Google Scholar]
  9. Kleemola SRM, Karjalainen JE, Ràty RKH. Rapid diagnosis of Mycoplasma pneumoniae infection: clinical evaluation of a commercial probe test. J Infect Dis 1990; 162:70–75
    [Google Scholar]
  10. Saiki RK, Scharf S, Faloona F. et al. Enzymatic amplification of β-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science 1985; 230:1350–1354
    [Google Scholar]
  11. 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; 27:2492–2496
    [Google Scholar]
  12. Jensen JS, Sondergàrd-Andersen J, Uldum SA, Lind K. Detection of Mycoplasma pneumoniae in simulated clinical samples by polymerase chain reaction. APMIS 1989; 97:1046–1048
    [Google Scholar]
  13. Chanock RM, Hayflick L, Barile MF. Growth on artificial medium of an agent associated with atypical pneumonia and its identification as a PPLO. Proc Natl Acad Sci USA 1962; 48:41–49
    [Google Scholar]
  14. Kraybill WH, Crawford YE. A selective medium and color test for Mycoplasma pneumoniae. Proc Soc Exp Biol Med 1965; 118:965–970
    [Google Scholar]
  15. Smith CB, Chanock RM, Friedewald WT. et al. Mycoplasma pneumoniae infections in volunteers. Ann NY Acad Sci 1967; 143:471–483
    [Google Scholar]
  16. Stauffer GV, Plamann MD, Stauffer LT. Construction and expression of hybrid plasmids containing the Escherichia coli glr A genes. Gene 1981; 14:63–72
    [Google Scholar]
  17. Saiki RK, Gelfand DH, Stoffel S. et al. Primer-directed enzymatic amplification of DNA with a thermostable DN A polymerase. Science 1988; 239:487–491
    [Google Scholar]
  18. Weisburg WG, Tully JG, Rose DL. et al. A phylogenetic analysis of the mycoplasmas: basis for their classification. J Bacterial 1989; 171:6455–6467
    [Google Scholar]
  19. Kai M, Kamiya S, Sawamura S, Yamamoto T, Ozawa A. Simplified method for confirmation of PCR products. Nucleic Acids Res 1991; 19:4562
    [Google Scholar]
  20. Noller HF, Woese CR. Secondary structure of 16S ribosomal RNA. Science 1981; 212:403–411
    [Google Scholar]
  21. Gray MW, Sankoff D, Cedergren RJ. On the evolutionary descent of organisms and organelles: a global phytogeny based on a highly conserved structural core in small subunit ribosomal RNA. Nucleic Acids Res 1984; 12:5837–5852
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/00222615-38-3-166
Loading
/content/journal/jmm/10.1099/00222615-38-3-166
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error