Skip to main content
PMC home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1996 Apr;34(4):785–791. doi: 10.1128/jcm.34.4.785-791.1996

Diagnosis of contagious caprine pleuropneumonia by detection and identification of Mycoplasma capricolum subsp. capripneumoniae by PCR and restriction enzyme analysis.

G Bölske 1, J G Mattsson 1, C R Bascuñana 1, K Bergström 1, H Wesonga 1, K E Johansson 1
PMCID: PMC228893  PMID: 8815084

Abstract

Contagious caprine pleuropneumonia (CCPP), one of the most serious and dramatic diseases of goats, is caused by Mycoplasma capricolum subsp. capripneumoniae (M. capripneumoniae). This organism is very difficult to isolate and to correctly identify. In a previous report we described a method for the rapid detection and identification of M. capripneumoniae. This method is based on a PCR system by which a segment of the 16S rRNA gene from all mycoplasmas of the M. mycoides cluster can be amplified. The PCR product is then analyzed by restriction enzyme cleavage for the identification of M. capripneumoniae DNA. This system has now been further evaluated with respect to specificity and diagnostic efficacy for the identification and direct detection of the organism in clinical material. Identification by restriction enzyme analysis of amplified DNA from mycoplasmas of the M. mycoides cluster was verified for 55 strains, among which were 15 strains of M. capripneumoniae. The PCR was applied to clinical samples from the nose, ear, pharynx, pleural fluid, and lung tissue containing M. capripneumoniae or other mycoplasmas. As expected, mycoplasmas belonging to the M. mycoides cluster could be detected by the PCR. Restriction enzyme analysis of the PCR products could then be applied for the identification of M. capripneumoniae. Clinical samples and cultures containing M. capripneumoniae were dried on filter paper, to try an easier sample transport method, and were tested by PCR. M. capripneumoniae DNA could be detected in the dried specimens, but the sensitivity of the PCR test was reduced.

Full Text

The Full Text of this article is available as a PDF (230.4 KB).

Selected References

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

  1. Bascuñana C. R., Mattsson J. G., Bölske G., Johansson K. E. Characterization of the 16S rRNA genes from Mycoplasma sp. strain F38 and development of an identification system based on PCR. J Bacteriol. 1994 May;176(9):2577–2586. doi: 10.1128/jb.176.9.2577-2586.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Belton D., Leach R. H., Mitchelmore D. L., Rurangirwa F. R. Serological specificity of a monoclonal antibody to Mycoplasma capricolum strain F38, the agent of contagious caprine pleuropneumonia. Vet Rec. 1994 Jun 18;134(25):643–646. doi: 10.1136/vr.134.25.643. [DOI] [PubMed] [Google Scholar]
  3. Black F. T. Modifications of the growth inhibition test and its application to human T-mycoplasmas. Appl Microbiol. 1973 Apr;25(4):528–533. doi: 10.1128/am.25.4.528-533.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bonnet F., Saillard C., Bové J. M., Leach R. H., Rose D. L., Cottew G. S., Tully J. G. DNA relatedness between field isolates of Mycoplasma F38 group, the agent of contagious caprine pleuropneumonia, and strains of Mycoplasma capricolum. Int J Syst Bacteriol. 1993 Jul;43(3):597–602. doi: 10.1099/00207713-43-3-597. [DOI] [PubMed] [Google Scholar]
  5. Bölske G., Johansson K. E., Heinonen R., Panvuga P. A., Twinamasiko E. Contagious caprine pleuropneumonia in Uganda and isolation of Mycoplasma capricolum subspecies capripneumoniae from goats and sheep. Vet Rec. 1995 Dec 2;137(23):594–594. [PubMed] [Google Scholar]
  6. Bölske G., Msami H. M., Gunnarsson A., Kapaga A. M., Loomu P. M. Contagious bovine pleuropneumonia in northern Tanzania, culture confirmation and serological studies. Trop Anim Health Prod. 1995 Nov;27(4):193–201. doi: 10.1007/BF02250690. [DOI] [PubMed] [Google Scholar]
  7. Bölske G., Msami H., Humlesjö N. E., Ernø H., Jönsson L. Mycoplasma capricolum in an outbreak of polyarthritis and pneumonia in goats. Acta Vet Scand. 1988;29(3-4):331–338. doi: 10.1186/BF03548626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bölske G. Survey of Mycoplasma infections in cell cultures and a comparison of detection methods. Zentralbl Bakteriol Mikrobiol Hyg A. 1988 Nov;269(3):331–340. doi: 10.1016/s0176-6724(88)80176-7. [DOI] [PubMed] [Google Scholar]
  9. DaMassa A. J., Tully J. G., Rose D. L., Pitcher D., Leach R. H., Cottew G. S. Mycoplasma auris sp. nov., Mycoplasma cottewii sp. nov., and Mycoplasma yeatsii sp. nov., new sterol-requiring mollicutes from the external ear canals of goats. Int J Syst Bacteriol. 1994 Jul;44(3):479–484. doi: 10.1099/00207713-44-3-479. [DOI] [PubMed] [Google Scholar]
  10. DaMassa A. J., Wakenell P. S., Brooks D. L. Mycoplasmas of goats and sheep. J Vet Diagn Invest. 1992 Jan;4(1):101–113. doi: 10.1177/104063879200400126. [DOI] [PubMed] [Google Scholar]
  11. Harbi M. S., El Tahir M. S., Macowan K. J., Nayil A. A. Mycoplasma strain F38 and contagious caprine pleuropneumonia in the Sudan. Vet Rec. 1981 Mar 21;108(12):261–261. doi: 10.1136/vr.108.12.261. [DOI] [PubMed] [Google Scholar]
  12. Jones G. E., Rae A. G., Holmes R. G., Lister S. A., Jones J. M., Grater G. S., Richards N. Isolation of exotic mycoplasmas from sheep in England. Vet Rec. 1983 Dec 3;113(23):540–540. [PubMed] [Google Scholar]
  13. Jones G. E., Wood A. R. Microbiological and serological studies on caprine pneumonias in Oman. Res Vet Sci. 1988 Jan;44(1):125–131. [PubMed] [Google Scholar]
  14. Leach R. H. Comparative studies of mycoplasma of bovine origin. Ann N Y Acad Sci. 1967 Jul 28;143(1):305–316. doi: 10.1111/j.1749-6632.1967.tb27670.x. [DOI] [PubMed] [Google Scholar]
  15. Leach R. H., Ernø H., MacOwan K. J. Proposal for designation of F38-type caprine mycoplasmas as Mycoplasma capricolum subsp. capripneumoniae subsp. nov. and consequent obligatory relegation of strains currently classified as M. capricolum (Tully, Barile, Edward, Theodore, and Ernø 1974) to an additional new subspecies, M. capricolum subsp. capricolum subsp. nov. Int J Syst Bacteriol. 1993 Jul;43(3):603–605. doi: 10.1099/00207713-43-3-603. [DOI] [PubMed] [Google Scholar]
  16. Lefevre P. C., Breard A., Alfarouk I., Buron S. Mycoplasma species F 38 isolated in Chad. Vet Rec. 1987 Dec 12;121(24):575–576. [PubMed] [Google Scholar]
  17. Litamoi J. K., Wanyangu S. W., Simam P. K. Isolation of Mycoplasma biotype F38 from sheep in Kenya. Trop Anim Health Prod. 1990 Nov;22(4):260–262. doi: 10.1007/BF02240409. [DOI] [PubMed] [Google Scholar]
  18. MacOwan K. J., Minette J. E. A mycoplasma from acute contagious caprine pleuropneumonia in Kenya. Trop Anim Health Prod. 1976 May;8(2):91–95. doi: 10.1007/BF02383376. [DOI] [PubMed] [Google Scholar]
  19. McMartin D. A., MacOwan K. J., Swift L. L. A century of classical contagious caprine pleuropneumonia: from original description to aetiology. Br Vet J. 1980 Sep-Oct;136(5):507–515. doi: 10.1016/s0007-1935(17)32196-6. [DOI] [PubMed] [Google Scholar]
  20. Olsson B., Bölske G., Bergström K., Johansson K. E. Analysis of caprine mycoplasmas and mycoplasma infections in goats using two-dimensional electrophoresis and immunoblotting. Electrophoresis. 1990 Oct;11(10):861–869. doi: 10.1002/elps.1150111016. [DOI] [PubMed] [Google Scholar]
  21. Perreau P., Breard A. La mycoplasmose caprine à M. capricolum. Comp Immunol Microbiol Infect Dis. 1979;2(1):87–97. doi: 10.1016/0147-9571(79)90062-6. [DOI] [PubMed] [Google Scholar]
  22. Perreau P., Breard A., Le Goff C. Infection expérimentale de la chèvre par les souches de mycoplasme de type F.38 (pleuropneumonie contagieuse caprine). Ann Microbiol (Paris) 1984 Jan-Feb;135A(1):119–124. [PubMed] [Google Scholar]
  23. Rosendal S., Black F. T. Direct and indirect immunofluorescence of unfixed and fixed Mycoplasma colonies. Acta Pathol Microbiol Scand B Microbiol Immunol. 1972;80(4):615–622. doi: 10.1111/j.1699-0463.1972.tb00186.x. [DOI] [PubMed] [Google Scholar]
  24. Rurangirwa F. R., McGuire T. C., Musoke A. J., Kibor A. Differentiation of F38 mycoplasmas causing contagious caprine pleuropneumonia with a growth-inhibiting monoclonal antibody. Infect Immun. 1987 Dec;55(12):3219–3220. doi: 10.1128/iai.55.12.3219-3220.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Thiaucourt F., Breard A., Lefèvre P. C., Mebratu G. Y. Contagious caprine pleuropneumonia in Ethiopia. Vet Rec. 1992 Dec 19;131(25-26):585–585. [PubMed] [Google Scholar]
  26. Thiaucourt F., Bölske G., Libeau G., Le Goff C., Lefèvre P. C. The use of monoclonal antibodies in the diagnosis of contagious caprine pleuropneumonia (CCPP). Vet Microbiol. 1994 Aug 1;41(3):191–203. doi: 10.1016/0378-1135(94)90100-7. [DOI] [PubMed] [Google Scholar]
  27. Thiaucourt F., Guérin C., Mady V., Lefèvre P. C. Diagnostic de la pleuropneumonie contagieuse caprine: améliorations récentes. Rev Sci Tech. 1992 Sep;11(3):859–865. [PubMed] [Google Scholar]
  28. Tully J. G., Barile M. F., Edward D. G., Theodore T. S., Erno H. Characterization of some caprine mycoplasmas, with proposals for new species, mycoplasma capricolum and mycoplasma putrefaciens. J Gen Microbiol. 1974 Nov;85(1):102–120. doi: 10.1099/00221287-85-1-102. [DOI] [PubMed] [Google Scholar]
  29. al-Aubaidi J. M., Fabricant J. Characterization and classification of bovine mycoplasma. Cornell Vet. 1971 Jul;61(3):490–518. [PubMed] [Google Scholar]

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

RESOURCES