1887

Abstract

, the aetiological agent of the plague, causes sporadic disease in endemic areas of the world and is classified as a National Institute of Allergy and Infectious Diseases Category A Priority Pathogen because of its potential to be used as a bioweapon. Health departments, hospitals and government agencies need the ability to rapidly identify and characterize cultured isolates of this bacterium. Assays have been developed to perform this function; however, they are limited in their ability to distinguish from . This report describes the creation of a real-time PCR assay using Taqman probes that exclusively identifies using a unique target sequence of the gene on the chromosome. As with other PCR assays, three major genes located on each of the three virulence plasmids were included: on pCD1, on pMT1 and on pPCP1. The quadruplex assay was validated on a collection of 192 isolates and 52 near-neighbour isolates. It was discovered that only 72 % of natural plague isolates from the states of New Mexico and Utah harboured all three virulence plasmids. This quadruplex assay proved to be 100 % successful in differentiating from all near neighbours tested and was able to reveal which of the three virulence plasmids a particular isolate possessed.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.47485-0
2008-03-01
2021-04-22
Loading full text...

Full text loading...

/deliver/fulltext/jmm/57/3/324.html?itemId=/content/journal/jmm/10.1099/jmm.0.47485-0&mimeType=html&fmt=ahah

References

  1. Achtman M., Zurth K., Morelli G., Torrea G., Guiyoule A., Carniel E. 1999; Yersinia pestis , the cause of plague, is a recently emerged clone of Yersinia pseudotuberculosis . Proc Natl Acad Sci U S A 96:14043–14048 [CrossRef]
    [Google Scholar]
  2. Brubaker R. R. 2004; The recent emergence of plague: a process of felonious evolution. Microb Ecol 47:293–299
    [Google Scholar]
  3. Brubaker R. R., Beesley E. D., Surgalla M. J. 1965; Pasteurella pestis : role of pesticin I and iron in experimental plague. Science 149:422–424 [CrossRef]
    [Google Scholar]
  4. Campbell J., Lowe J., Walz S., Ezzell J. 1993; Rapid and specific identification of Yersinia pestis by using a nested polymerase chain reaction procedure. J Clin Microbiol 31:758–759
    [Google Scholar]
  5. de Almeida A. M., Guiyoule A., Guilvout I., Iteman I., Baranton G., Carniel E. 1993; Chromosomal irp2 gene in Yersinia : distribution, expression, deletion and impact on virulence. Microb Pathog 14:9–21 [CrossRef]
    [Google Scholar]
  6. Devdariani Z. L., Verenkov M. S., Feodorova V. A., Solodovnicov N. S., Belov L. G. 1993; Identification of Yersinia pestis with varied plasmid composition using monoclonal and polyclonal fluorescent immunoglobulins. FEMS Immunol Med Microbiol 6:31–35 [CrossRef]
    [Google Scholar]
  7. Drozdov I. G., Anisimov A. P., Samoilova S. V., Yezhov I. N., Yeremin S. A., Karlyshev A. V., Krasilnikova V. M., Kravchenko V. I. 1995; Virulent non-capsulate Yersinia pestis variants constructed by insertion mutagenesis. J Med Microbiol 42:264–268 [CrossRef]
    [Google Scholar]
  8. Engelthaler D. M., Gage K. L., Montenieri J. A., Chu M., Carter L. G. 1999; PCR detection of Yersinia pestis in fleas: comparison with mouse inoculation. J Clin Microbiol 37:1980–1984
    [Google Scholar]
  9. Feodorova V. A., Devdariani Z. L. 2000; Development, characterisation and diagnostic application of monoclonal antibodies against Yersinia pestis fibrinolysin and coagulase. J Med Microbiol 49:261–269
    [Google Scholar]
  10. Ferber D. M., Brubaker R. R. 1981; Plasmids in Yersinia pestis . Infect Immun 31:839–841
    [Google Scholar]
  11. Filippov A. A., Solodovnikov N. S., Kookleva L. M., Protsenko O. A. 1990; Plasmid content in Yersinia pestis strains of different origin. FEMS Microbiol Lett 55:45–48
    [Google Scholar]
  12. Higgins J. A., Ezzell J., Hinnebusch B. J., Shipley M., Henchal E. A., Ibrahim M. S. 1998; 5′ Nuclease PCR assay to detect Yersinia pestis . J Clin Microbiol 36:2284–2288
    [Google Scholar]
  13. Hinnebusch J., Schwan T. G. 1993; New method for plague surveillance using polymerase chain reaction to detect Yersinia pestis in fleas. J Clin Microbiol 31:1511–1514
    [Google Scholar]
  14. Hu P., Elliott J., McCready P., Skowronski E., Garnes J., Kobayashi A., Brubaker R. R., Garcia E. 1998; Structural organization of virulence-associated plasmids of Yersinia pestis . J Bacteriol 180:5192–5202
    [Google Scholar]
  15. Iqbal S. S., Chambers J. P., Goode M. T., Valdes J. J., Brubaker R. R. 2000; Detection of Yersinia pestis by pesticin fluorogenic probe-coupled PCR. Mol Cell Probes 14:109–114 [CrossRef]
    [Google Scholar]
  16. Khushiramani R., Shukla J., Tuteja U., Batra H. V. 2006; Yersinia outer-membrane protein B (YopB): a tool for identification of Yersinia pestis isolates. J Med Microbiol 55:467–469 [CrossRef]
    [Google Scholar]
  17. Leal N. C., Almeida A. M. 1999; Diagnosis of plague and identification of virulence markers in Yersinia pestis by multiplex-PCR. Rev Inst Med Trop Sao Paulo 41:339–342 [CrossRef]
    [Google Scholar]
  18. Leal N. C., Abath F. G., Alves L. C., Almeida A. M. 1996; A simple PCR-based procedure for plague diagnosis. Rev Inst Med Trop Sao Paulo 38:371–373 [CrossRef]
    [Google Scholar]
  19. Leal-Balbino T. C., Leal N. C., Lopes C. V., Almeida A. M. 2004; Differences in the stability of the plasmids of Yersinia pestis cultures in vitro: impact on virulence. Mem Inst Oswaldo Cruz 99:727–732 [CrossRef]
    [Google Scholar]
  20. Li H., Bhaduri S., Magee W. E. 1998; Maximizing plasmid stability and production of released proteins in Yersinia enterocolitica . Appl Environ Microbiol 64:1812–1815
    [Google Scholar]
  21. Loiez C., Herwegh S., Wallet F., Armand S., Guinet F., Courcol R. J. 2003; Detection of Yersinia pestis in sputum by real-time PCR. J Clin Microbiol 41:4873–4875 [CrossRef]
    [Google Scholar]
  22. McAvin J. C., McConathy M. A., Rohrer A. J., Huff W. B., Barnes W. J., Lohman K. L. 2003; A real-time fluorescence polymerase chain reaction assay for the identification of Yersinia pestis using a field-deployable thermocycler. Mil Med 168:852–855
    [Google Scholar]
  23. McDonough K. A., Schwan T. G., Thomas R. E., Falkow S. 1988; Identification of a Yersinia pestis -specific DNA probe with potential for use in plague surveillance. J Clin Microbiol 26:2515–2519
    [Google Scholar]
  24. Moore R. L., Brubaker R. R. 1975; Hybridization of deoxyribonucleotide sequences of Yersinia enterocolitica and other selected members of Enterobacteriaceae . Int J Syst Bacteriol 25:336–339 [CrossRef]
    [Google Scholar]
  25. Neubauer H., Meyer H., Prior J., Aleksic S., Hensel A., Splettstösser W. 2000; A combination of different polymerase chain reaction (PCR) assays for the presumptive identification of Yersinia pestis . J Vet Med B Infect Dis Vet Public Health 47:573–580 [CrossRef]
    [Google Scholar]
  26. Norkina O. V., Kulichenko A. N., Gintsburg A. L., Tuchkov I. V., Popov Yu. A., Aksenov M. U., Drosdov I. G. 1994; Development of a diagnostic test for Yersinia pestis by the polymerase chain reaction. J Appl Bacteriol 76:240–245 [CrossRef]
    [Google Scholar]
  27. Parkhill J., Wren B. W., Thomson N. R., Titball R. W., Holden M. T., Prentice M. B., Sebaihia M., James K. D., Churcher C. other authors 2001; Genome sequence of Yersinia pestis , the causative agent of plague. Nature 413:523–527 [CrossRef]
    [Google Scholar]
  28. Parmenter R. R., Yadav E. P., Parmenter C. A., Ettestad P., Gage K. L. 1999; Incidence of plague associated with increased winter–spring precipitation in New Mexico. Am J Trop Med Hyg 61:814–821
    [Google Scholar]
  29. Perry R. D., Fetherston J. D. 1997; Yersinia pestis – etiologic agent of plague. Clin Microbiol Rev 10:35–66
    [Google Scholar]
  30. Portnoy D. A., Falkow S. 1981; Virulence-associated plasmids from Yersinia enterocolitica and Yersinia pestis . J Bacteriol 148:877–883
    [Google Scholar]
  31. Sebbane F., Jarrett C. O., Gardner D., Long D., Hinnebusch B. J. 2006; Role of the Yersinia pestis plasminogen activator in the incidence of distinct septicemic and bubonic forms of flea-borne plague. Proc Natl Acad Sci U S A 103:5526–5530 [CrossRef]
    [Google Scholar]
  32. Sodeinde O. A., Goguen J. D. 1988; Genetic analysis of the 9.5-kilobase virulence plasmid of Yersinia pestis . Infect Immun 56:2743–2748
    [Google Scholar]
  33. Thoerner P., Bin Kingombe C. I., Bögli-Stuber K., Bissig-Choisat B., Wassenaar T. M., Frey J., Jemmi T. 2003; PCR detection of virulence genes in Yersinia enterocolitica and Yersinia pseudotuberculosis and investigation of virulence gene distribution. Appl Environ Microbiol 69:1810–1816 [CrossRef]
    [Google Scholar]
  34. Thomson N. R., Howard S., Wren B. W., Holden M. T., Crossman L., Challis G. L., Churcher C., Mungall K., Brooks K. other authors 2006; The complete genome sequence and comparative genome analysis of the high pathogenicity Yersinia enterocolitica strain 8081. PLoS Genet 2:e206 [CrossRef]
    [Google Scholar]
  35. Tomaso H., Reisinger E. C., Al Dahouk S., Frangoulidis D., Rakin A., Landt O., Neubauer H. 2003; Rapid detection of Yersinia pestis with multiplex real-time PCR assays using fluorescent hybridisation probes. FEMS Immunol Med Microbiol 38:117–126 [CrossRef]
    [Google Scholar]
  36. Tsukano H., Itoh K., Suzuki S., Watanabe H. 1996; Detection and identification of Yersinia pestis by polymerase chain reaction (PCR) using multiplex primers. Microbiol Immunol 40:773–775 [CrossRef]
    [Google Scholar]
  37. Woron A. M., Nazarian E. J., Egan C., McDonough K. A., Cirino N. M., Limberger R. J., Musser K. A. 2006; Development and evaluation of a 4-target multiplex real-time polymerase chain reaction assay for the detection and characterization of Yersinia pestis . Diagn Microbiol Infect Dis 56:261–268 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.47485-0
Loading
/content/journal/jmm/10.1099/jmm.0.47485-0
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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