Skip to main content
SpringerLink
Log in

Detection and Identification of Mycoplasmas with Diagnostic DNA Probes Complementary to Ribosomal RNA

  • Chapter
Rapid Diagnosis of Mycoplasmas

Part of the book series: Federation of European Microbilogical Societies Symposium Series ((FEMS,volume 62))

Abstract

Hybridization is the reaction by which two single stranded and complementary nucleic acid molecules can form a duplex molecule by base pairing through hydrogen bonds. This reaction has been known for more than 30 years (Hall and Spiegelman, 1961) and proved very useful in research but has, not until recently, been utilized in diagnostic microbiology (for a review, see Tenover, 1988). Nucleic acid molecules used in the hybridization reaction for a diagnostic or a detective purpose are referred to as nucleic acid (DNA or RNA) probes, gene probes or simply probes. The first generation of probes was composed of labeled pieces of whole genomic DNA and these probes gave a comparatively low specificity, but have been extensively used to study phylogenetic relatedness of bacteria in so-called DNA hybridization experiments. The second generation of probes are those which are composed of labeled restriction endonuclease fragments and produced by molecular cloning. These probes represent a more well-defined system and give, in general, a higher specificity. The third generation of probes consists of short oligonucleotides (15–50 nucleotides in length) and they are produced by chemical oligonucleotide synthesis from available sequence information. Oligonucleotide probes give the highest possible specificity and diagnostic methods based on oligonucleotide probes represent extremely well-defined and reproducible systems. As an example, if the target molecule for an oligonucleotide probe of 15–20 nucleotides in length has one single mismatched base pair of the unstable type (Ikuta et al., 1987) close to the middle of the duplex molecule, the stringency can be selected to avoid cross-hybridization with this particular target.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
eBook
USD 16.99
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 16.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  • Blanchard, A., Gautier, M. and Mayau, V. (1991) Detection and identification of mycoplasmas by amplification of rDNA. FEMS Microbiol. Lett. 81:37–42.

    CAS  Google Scholar 

  • Chen, X. and Finch, L. R. (1989) Novel arrangement of rRNA genes in Mycoplasma gallisepticum: Separation of the 16S rRNA gene of one set from the 23S and 5S genes. J. Bacteriol. 171:2876–2878.

    PubMed  CAS  Google Scholar 

  • De Rijk, P., Neefs, J.-M., Van de Peer, Y. and De Wachter, R. (1992) Compilation of small subunit RNA sequences. Nucleic Acids Res. Suppl. 20:2075–2089.

    Article  Google Scholar 

  • Devereux, J., Haeberli, P. and Smithies, O. (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 12:387–395.

    Article  PubMed  CAS  Google Scholar 

  • Fernández, C., Mattsson, J. G., Bölske, G., Levisohn, S. and Johansson, K.-E. (1993) Species-specific oligonucleotide probes complementary to 16S-rRNA of Mycoplasma gallisepticum and Mycoplasma synoviae. Res. Vet. Sci. (in press).

    Google Scholar 

  • Frydenberg, J. and Christiansen, C. (1985) The sequence of 16S rRNA from mycoplasma strain PG50. DNA 4:127–137.

    Article  PubMed  CAS  Google Scholar 

  • Futo, S., Soto, Y., Mitsuse, S. and Mori, Y. (1992) Detection of Mycoplasma hyopneumoniae by using rRNA-oligonucleotide hybridization. J. Clin. Microbiol. 30:1509–1513.

    PubMed  CAS  Google Scholar 

  • Göbel, U.B. and Stanbridge, E. J. (1984) Cloned mycoplasma ribosomal RNA genes for the detection of mycoplasma contamination in tissue cultures. Science 226:1211–1213.

    Article  PubMed  Google Scholar 

  • Göbel, U., Maas, R., Haun, G., Vinga-Martins, C. and Stanbridge, E. J. (1987a) Synthetic oligonucleotide probes complementary to rRNA for group-and species-specific detection of mycoplasmas. Israel J. Med. Sci. 23:742–746.

    PubMed  Google Scholar 

  • Göbel, U.B., Geiser, A. and Stanbridge, E. J. (1987b) Oligonucleotide probes complementary to variable regions of ribosomal RNA discriminate between Mycoplasma species. J. Gen. Microbiol. 133:1969–1974.

    PubMed  Google Scholar 

  • Grau, O., Laigret, F., Carie, P., Tully, J. G., Rose, D. and Bové, J. M. (1991) Identification of a plant-derived Mollicute as a strain of an avian pathogen, Mycoplasma iowae, and its implications for Mollicute taxonomy. Int. J. Syst. Bacteriol. 41:473–478.

    Article  PubMed  CAS  Google Scholar 

  • Gray, M. W., Sankoff, D. and Cedergren, R. J. (1984) On the evolutionary descent of organisms and organelles: a global phylogeny based on a highly conserved structural core in small subunit ribosomal RNA. Nucleic Acids Res. 12:5837–5852.

    Article  PubMed  CAS  Google Scholar 

  • Hall, B. D. and Spiegelman, S. (1961) Sequence complementarity of T2-DNA and T2-specific RNA. Proc. Natl. Acad. Sci. 47:137–146.

    Article  PubMed  CAS  Google Scholar 

  • Ikuta, S., Takagi, K, Wallace, R. B. and Itakura, K. (1987) Dissociation kinetics of 19 base paired oligonucleotide-DNA duplexes containing different single mismatched base pairs. Nucleic Acids Res. 15:797–811.

    Article  PubMed  CAS  Google Scholar 

  • Iwami, M., Muto, A., Yamao, F. and Osawa, S. (1984) Nucleotide sequence of the rrnB 16S ribosomal RNA gene from Mycoplasma capricolum. Mol. Gen. Genet. 196, 317–322.

    Article  PubMed  CAS  Google Scholar 

  • Johansson, K.-E. and Bölske, G. (1989) Evaluation and practical aspects of the use of a commercial DNA probe for detection of mycoplasma infections in cell cultures. J. Biochem. Biophys. Methods 19:185–200.

    Article  PubMed  CAS  Google Scholar 

  • Johansson, K.-E., Johansson, I. and Göbel, U. B. (1990) Evaluation of different hybridization procedures for the detection of mycoplasma contamination in cell cultures. Mol. Cell. Probes 4:33–42.

    Article  PubMed  CAS  Google Scholar 

  • Johansson, K.-E., Mattsson, J.G., Jacobsson, K., Fernández, C., Bergström, K., Bölske, G., Wallgren, P. and Göbel, U. B. (1992) Specificity of oligonucleotide probes complementary to evolutionarily variable regions of 16S rRNA from Mycoplasma hyopneumoniae and Mycoplasma hyorhinis. Res. Vet. Sci. 52:195–204.

    Article  PubMed  CAS  Google Scholar 

  • Kwok, S., Kellogg, D. E., McKinney, N., Spasic, D., Goda, L., Levenson, C. and Sninsky, J. J. (1990) Effects of primer-template mismatches on the polymerase chain reaction: human immunodeficiency virus type 1 model studies. Nucleic Acids Res. 18:999–1005.

    Article  PubMed  CAS  Google Scholar 

  • Lane, D. J., Pace, B., Olsen, G. J., Stahl, D. A., Sogin, M. L. and Pace, N.R. (1985) Rapid determination of 16S ribosomal RNA sequences for phylogenetic analysis. Proc. Natl. Acad. Sci. USA 82:6955–6959.

    Article  PubMed  CAS  Google Scholar 

  • Maniloff, J. (1992) Phylogeny of Mycoplasmas, in “Mycoplasmas: Molecular Biology and Pathogenesis” (Maniloff, J., McElhaney, R. N., Finch, L. R. and Baseman, J. B., Eds.), pp. 549–559, ASM Press, Washington.

    Google Scholar 

  • Mattsson, J. G. and Johannsson, K.-E. (1993) Oligonucleotide probes complementary to 16S rRNA for rapid detection of mycoplasma contamination in cell cultures. FEMS Microbiol. Lett. 107:139–144.

    Article  PubMed  CAS  Google Scholar 

  • Mattsson, J. G., Gersdorf, H., Göbel, U.B. and Johansson, K.-E. (1991) Detection of Mycoplasma bovis and Mycoplasma agalactiae by oligonucleotide probes complementary to 16S rRNA. Mol. Cell. Probes 5:27–35.

    Article  PubMed  CAS  Google Scholar 

  • Mullis, K. B. and Faloona, F. A. (1987) Specific synthesis of DNA in vitro: a polymerase-catalyzed chain reaction. Methods Enzymol. 155:335–350.

    Article  PubMed  CAS  Google Scholar 

  • McGarrity, G. J. and Kotani, H. (1986) Detection of cell culture mycoplasmas by a genetic probe. Exp. Cell Res. 163:273–278.

    Article  PubMed  CAS  Google Scholar 

  • Nakagawa, T., Uemori, T., Asada, K., Kato, I. and Harasawa, R. (1992) Acholeplasma laidlawii has tRNA genes in the 16S-23S spacer of the rRNA operon. J. Bacteriol. 174:8163–8165.

    PubMed  CAS  Google Scholar 

  • Nur, I., Bové, J. M., Saillard, C., Rottem, S., Whitcomb, R. M. and Razin, S. (1986) DNA probes in detection of spiroplasmas and mycoplasma-like organisms in plants and insects. FEMS Microbiol. Lett. 35:157–162.

    Article  CAS  Google Scholar 

  • Nur, I., Reinhartz, A., Hyman, H. C., Razin, S., Herzberg, M. (1989) Chemiprobe™, a nonradioactive system for labeling nucleic acid. Principles and applications. Ann. Biol. Clin. 47:601–606.

    CAS  Google Scholar 

  • Olsen, G. J. and Woese, C.R. (1993) Ribosomal RNA: a key to phylogeny. FASEB J. 7:113–123.

    PubMed  CAS  Google Scholar 

  • Olsen, G. J., Overbeek, R., Larsen, N., Marsh, T. L., McCaughey, M. J., Maciukenas, M. A., Kuan, W.-M., Macke, T. J., Xing, Y. and Woese, C. R. (1992) The ribosomal database project. Nucleic Acids Res. Suppl. 20:2120–2199.

    Google Scholar 

  • Pettersson, B., Johansson, K.-E. and Uhlén, M. (1993) Direct automated solid phase sequencing in an in vitro amplified segment of 16S rDNA from bacteria, in: “Proceedings of the 6th European Congress on Biotechnology” (Alberghina, L., Frontali, L. and Sensi, P., Eds.). Elsevier, Amsterdam (in press).

    Google Scholar 

  • Razin, S. (1985) Molecular biology and genetics of mycoplasmas (Mollicutes). Microbiol. Rev. 49:419–455.

    PubMed  CAS  Google Scholar 

  • Razin, S. (1989) Molecular approach to mycoplasma phylogeny, in: “The Mycoplasmas, Vol. V. Spiroplasmas, Acholeplasmas and Mycoplasmas of Plants and Arthropods” (Whitcomb, R. F. and Tully, J. G. Eds.), pp. 33–69. Academic Press, New York.

    Google Scholar 

  • Razin, S., Gross, M., Wormser, M., Pollack, Y. and Glaser, G. (1984) Detection of mycoplasmas infecting cell cultures by DNA hybridization. In Vitro 20:404–408.

    Article  PubMed  CAS  Google Scholar 

  • Robertson, J. A., Vekris, A., Bébéar, C. and Stemke, G. W. (1993) Polymerase chain reaction using 16S rRNA gene sequences distinguishes the two biovars of Ureaplasma urealyticum. J. Clin. Microbiol. 31:824–830.

    PubMed  CAS  Google Scholar 

  • Shaw, S. B. (1989) DNA probes for the detection of Legionella species, Mycoplasma pneumoniae, and members of the Mycobacterium tuberculosis complex, in: “DNA Probes for Infectious Diseases” (Tenover, F. C., Ed.), pp. 101–117. CRC Press, Boca Raton.

    Google Scholar 

  • Sogin, M. L. (1990) Amplification of ribosomal RNA genes for molecular evolution studies, in: “PCR Protocols, a Guide to Methods and Applications” (Innis, M. A., Gelfund, D. H., Sninsky, J. J. and White, T. J., Eds.), pp. 307–317. Academic Press, New York.

    Google Scholar 

  • Stanbridge, E. J. (1976) A reevaluation of the role of mycoplasmas in human disease. Annu. Rev. Microbiol. 30:169–187.

    Article  PubMed  CAS  Google Scholar 

  • Stemke, G.W., Laigret, F., Grau, O. and Bové, J. M. (1992) Phylogenetic relationships of three porcine mycoplasmas, Mycoplasma hyopneumoniae, Mycoplasma flocculate, and Mycoplasma hyorhinis, and complete 16S rRNA sequences of M. flocculare. Int. J. Syst. Bacteriol. 42:220–225.

    Article  PubMed  CAS  Google Scholar 

  • Taschke, C., Klinkert, M.-Q., Wolters, J. and Herrmann, R. (1986) Organization of the ribosomal RNA genes in Mycoplasma hyopneumoniae: the 5S rRNA gene is separated from the 16S and 23S rRNA genes. Mol. Gen. Genet. 205:428–433.

    Article  PubMed  CAS  Google Scholar 

  • Taschke, C., Ruland, K. and Herrmann, R. (1987) Nucleotide sequence of the 16S rRNA of Mycoplasma hyopneumoniae. Nucleic Acids Res. 15:3918.

    Article  PubMed  CAS  Google Scholar 

  • Tenover, F. C. (1988) Diagnostic deoxyribonucleic acid probes for infectious diseases. Clin. Microbiol. Rev. 1:82–101.

    PubMed  CAS  Google Scholar 

  • Van Kuppeveld, F. J. M., van der Logt, J. T. M., Angulo, A. F., van Zoest, M. J., Quint, W. G.V., Niesters, H. G. M., Galama, J.M.D. and Melchers, W.J.G. (1992) Genus-and species-specific identification of mycoplasmas by 16S rRNA amplification. Appl. Environ. Microbiol. 58:2606–2615.

    PubMed  Google Scholar 

  • Wahlberg, J., Lundeberg, J., Hultman, T. and Uhlén, M. (1990) General colorimetric method for DNA diagnostics allowing direct solid-phase genomic sequencing of the positive samples. Proc. Natl. Acad. Sci. USA 87:6569–6573.

    Article  PubMed  CAS  Google Scholar 

  • Wahlberg, J., Holmberg, A., Berg, S., Hultman, T. and Uhlén, M. (1992) Automated magnetic preparation of DNA templates for solid phase sequencing. Electrophoresis 13:547–551.

    Article  PubMed  CAS  Google Scholar 

  • Weisburg, W. G., TuUy, J. G., Rose, D. L., Petzel, J. P., Oyaizu, H., Yang, D., Mandelco, L., Sechrest, J., Lawrence, T. G., Van Etten, J., Maniloff, J. and Woese, C.R. (1989) A phylogenetic analysis of the mycoplasmas: Basis for their classification. J. Bacteriol. 171:6455–6467.

    PubMed  CAS  Google Scholar 

  • Weisburg, W. G., Barns, S. M., Pelletier, D. A. and Lane, D. J. (1991) 16S ribosomal amplification for phylogenetic studies. J. Bacteriol. 173:697–703.

    PubMed  CAS  Google Scholar 

  • Woese, C. R. (1987) Bacterial evolution. Microbiol. Rev. 51:221–271.

    PubMed  CAS  Google Scholar 

  • Woese, C. R., Gutell, R., Gupta, R. and Noller, H. F. (1983) Detailed analysis of the higher-order structure of 16S-like ribosomal ribonucleic acids. Microbiol. Rev. 47:621–669.

    PubMed  CAS  Google Scholar 

  • Woese, C. R., Stackebrandt, E. and Ludwig, W. (1985) What are mycoplasmas: The relationship of tempo and mode in bacterial evolution. J. Mol. Evol. 21:305–316.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. The National Veterinary Institute, P.O. Box 7073, S-750 07, Uppsala, Sweden

    Karl-Erik Johansson

Authors
  1. Karl-Erik Johansson
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

Copyright information

© 1993 Springer Science+Business Media New York

About this chapter

Cite this chapter

Johansson, KE. (1993). Detection and Identification of Mycoplasmas with Diagnostic DNA Probes Complementary to Ribosomal RNA. In: Rapid Diagnosis of Mycoplasmas. Federation of European Microbilogical Societies Symposium Series, vol 62. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-2478-6_11

Download citation

  • DOI: https://doi.org/10.1007/978-1-4615-2478-6_11

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4613-6056-8

  • Online ISBN: 978-1-4615-2478-6

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics

Search

Navigation