Skip to main content
NCBI home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1994 Apr;32(4):1004–1010. doi: 10.1128/jcm.32.4.1004-1010.1994

Differentiation of toxigenic from nontoxigenic isolates of Pasteurella multocida by PCR.

S Nagai 1, S Someno 1, T Yagihashi 1
PMCID: PMC267170  PMID: 8027302

Abstract

A PCR assay was developed for the differentiation of toxigenic Pasteurella multocida subsp. multocida strains, the major etiologic agent for progressive atrophic rhinitis in pigs, from nontoxigenic strains. The PCR targeted a toxA gene encoding a 143-kDa dermonecrotic toxin that is considered to be the central etiologic factor in progressive atrophic rhinitis. toxA fragments were amplified from toxigenic P. multocida isolates but not from nontoxigenic isolates or other bacteria isolated from pigs. The sensitivity of the reaction was as low as 10 pg of chromosomal DNA from a toxigenic strain. The results obtained by PCR of the DNAs of 187 field isolates of P. multocida were consistent with those obtained by the guinea pig skin test and Western blot (immunoblot) analysis. Restriction fragment analysis of the PCR-amplified fragments from 67 field isolates and comparison of the DNA sequences of fragments from capsular serotype A and D strains suggest that the PCR-amplified region, which is considered to encode the major immunologic determinants of the toxin, would be the same among P. multocida strains. The PCR that we describe should be useful for the diagnosis and the etiologic survey of progressive atrophic rhinitis.

Full text

PDF
1004

Images in this article

1008Image
on p.1008
1009Image
on p.1009

Selected References

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

  1. Buys W. E., Smith H. E., Kamps A. M., Kamp E. M., Smits M. A. Sequence of the dermonecrotic toxin of Pasteurella multocida ssp. multocida. Nucleic Acids Res. 1990 May 11;18(9):2815–2816. doi: 10.1093/nar/18.9.2815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. CARTER G. R. Studies on Pasteurella multocida. I. A hemagglutination test for the identification of serological types. Am J Vet Res. 1955 Jul;16(60):481–484. [PubMed] [Google Scholar]
  3. Carter G. R., Rundell S. W. Identification of type A strains of P multocida using staphylococcal hyaluronidase. Vet Rec. 1975 Apr 12;96(15):343–343. doi: 10.1136/vr.96.15.343. [DOI] [PubMed] [Google Scholar]
  4. Dirks C., Schöos P., Schimmelpfennig H. Untersuchungen zur Atiologie der Rhinitis atrophicans des Schweines. Dtsch Tierarztl Wochenschr. 1973 Aug 1;80(15):342–contd. [PubMed] [Google Scholar]
  5. Foged N. T., Nielsen J. P., Pedersen K. B. Differentiation of toxigenic from nontoxigenic isolates of Pasteurella multocida by enzyme-linked immunosorbent assay. J Clin Microbiol. 1988 Jul;26(7):1419–1420. doi: 10.1128/jcm.26.7.1419-1420.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kamp E. M., ter Laak E. A., de Jong M. F. Atypical Pasteurella strains producing a toxin similar to the dermonecrotic toxin of Pasteurella multocida subspecies multocida. Vet Rec. 1990 Apr 28;126(17):434–437. [PubMed] [Google Scholar]
  7. Kamps A. M., Buys W. E., Kamp E. M., Smits M. A. Specificity of DNA probes for the detection of toxigenic Pasteurella multocida subsp. multocida strains. J Clin Microbiol. 1990 Aug;28(8):1858–1861. doi: 10.1128/jcm.28.8.1858-1861.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Magyar T., Rimler R. B. Detection and enumeration of toxin-producing Pasteurella multocida with a colony-blot assay. J Clin Microbiol. 1991 Jul;29(7):1328–1332. doi: 10.1128/jcm.29.7.1328-1332.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Nagai S., Takahashi K., Mitsui T., Yagihashi T. hlyIA-like sequence present in a genetic locus with low homology to appA in Actinobacillus pleuropneumoniae serotype 10. J Vet Med Sci. 1993 Oct;55(5):813–819. doi: 10.1292/jvms.55.813. [DOI] [PubMed] [Google Scholar]
  10. Pennings A. M., Storm P. K. A test in vero cell monolayers for toxin production by strains of Pasteurella multocida isolated from pigs suspected of having atrophic rhinitis. Vet Microbiol. 1984 Sep;9(5):503–508. doi: 10.1016/0378-1135(84)90071-3. [DOI] [PubMed] [Google Scholar]
  11. Petersen S. K., Foged N. T., Bording A., Nielsen J. P., Riemann H. K., Frandsen P. L. Recombinant derivatives of Pasteurella multocida toxin: candidates for a vaccine against progressive atrophic rhinitis. Infect Immun. 1991 Apr;59(4):1387–1393. doi: 10.1128/iai.59.4.1387-1393.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Petersen S. K. The complete nucleotide sequence of the Pasteurella multocida toxin gene and evidence for a transcriptional repressor, TxaR. Mol Microbiol. 1990 May;4(5):821–830. doi: 10.1111/j.1365-2958.1990.tb00652.x. [DOI] [PubMed] [Google Scholar]
  13. Rimler R. B., Brogden K. A. Pasteurella multocida isolated from rabbits and swine: serologic types and toxin production. Am J Vet Res. 1986 Apr;47(4):730–737. [PubMed] [Google Scholar]
  14. Rutter J. M., Luther P. D. Cell culture assay for toxigenic Pasteurella multocida from atrophic rhinitis of pigs. Vet Rec. 1984 Apr 21;114(16):393–396. doi: 10.1136/vr.114.16.393. [DOI] [PubMed] [Google Scholar]
  15. Rutter J. M. Virulence of Pasteurella multocida in atrophic rhinitis of gnotobiotic pigs infected with Bordetella bronchiseptica. Res Vet Sci. 1983 May;34(3):287–295. [PubMed] [Google Scholar]
  16. Saiki R. K., Scharf S., Faloona F., Mullis K. B., Horn G. T., Erlich H. A., Arnheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science. 1985 Dec 20;230(4732):1350–1354. doi: 10.1126/science.2999980. [DOI] [PubMed] [Google Scholar]
  17. Sawata A., Nakai T., Tuji M., Kume K. Dermonecrotic activity of Pasteurella multocida strains isolated from pigs in Japanese field. Nihon Juigaku Zasshi. 1984 Apr;46(2):141–148. doi: 10.1292/jvms1939.46.141. [DOI] [PubMed] [Google Scholar]
  18. Smith C. L., Econome J. G., Schutt A., Klco S., Cantor C. R. A physical map of the Escherichia coli K12 genome. Science. 1987 Jun 12;236(4807):1448–1453. doi: 10.1126/science.3296194. [DOI] [PubMed] [Google Scholar]
  19. de Jong M. F., Nielsen J. P. Definition of progressive atrophic rhinitis. Vet Rec. 1990 Jan 27;126(4):93–93. [PubMed] [Google Scholar]

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

RESOURCES