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Identification of encapsulated and non-encapsulated Yersinia pestis by immunofluorescence tests using polyclonal and monoclonal antibodies

Published online by Cambridge University Press:  15 May 2009

A. P. Phillips ,
B. C. Morris ,
D. Hall ,
M. Glenister  and
J. E. Williams
A. P. Phillips
Affiliation:
Chemical Defence Establishment, Porton Down, Salisbury, Wiltshire, SP4 OJQ, UK
B. C. Morris
Affiliation:
Chemical Defence Establishment, Porton Down, Salisbury, Wiltshire, SP4 OJQ, UK
D. Hall
Affiliation:
Chemical Defence Establishment, Porton Down, Salisbury, Wiltshire, SP4 OJQ, UK
M. Glenister
Affiliation:
Chemical Defence Establishment, Porton Down, Salisbury, Wiltshire, SP4 OJQ, UK
J. E. Williams
Affiliation:
Department of Hazardous Microorganisms, Walter Reed Army Institute of Research, Washington DC 20307–5100, USA
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Rabbit polyclonal hyperimmune antibodies to Yersinia pestis, and a mouse monoclonal antibody against the capsular antigen fraction 1 (F1) were compared in immunofluorescence (IF) tests. Fluorescent antibody conjugates were prepared from polyclonal antisera to four F1 positive Y. pestis strains; the conjugated antibody to strain A1122 gave the strongest IF staining of F1 positive and F1 negative Y. pestis strains. Indirect assays were rejected in favour of direct assays utilizing polyclonal and monoclonal reagents because the increased background staining reduced the effective contrast of bacterial visualisation. Polyclonal conjugates gave fairly homogeneous staining of Y. pestis bacterial populations, but in monoclonal assays a skew distribution of fluorescence intensity was observed, the majority of bacteria being poorly stained. The proportion of cells stained well by the monoclonal sufficed for easy identification of Y. pestis of the F1 positive phenotype however, and staining was not affected by washing the bacteria or treating them with formaldehyde. Y. pestis strains of the F1 positive genotype reacted with the monoclonal if bacteria were grown at 37 °C but not if the growth temperature was reduced to 25°C thus preventing capsule production. The polyclonal conjugate reacted with bacteria of these strains that had been grown at either temperature. Strains of F1 negative genotype grown at either temperature. Strains of F1 negative genotype grown at either temperature reacted with the polyclonal conjugate but not with the monoclonal. Cross reactions between the polyclonal reagents and Y. enterocolitica biovar 2, serovar O 8 could not be removed by selective absorption; however, the monoclonal antibody gave no cross reaction.

The F1 phenotypic status of bacterial preparations was verified by ELISA measurement of the fraction 1 antigen concentration. Antigen levels for F1 positive and F1 negative phenotypes differed by about three logs for suspensions of Y. pestis harvested from solid media.

The polyclonal and monoclonal direct IF tests applied to spleen and blood smears of laboratory mice infected with Y. pestis were able to differentiate between lethal infection with an F1 positive strain carrying all four classical virulence determinants, an F1 positive vaccine strain, and an F1 negative strain.

Type
Special Article
Information
Epidemiology & Infection , Volume 101 , Issue 1 , August 1988 , pp. 59 - 73
Copyright
Copyright © Cambridge University Press 1988

References

REFERENCES

Bahmanyar, M. & Cavanaugh, D. C. (1976). Plague Manual. Geneva: World Health Organisation.Google Scholar
Baker, E. E., Sommer, H., Foster, L. E., Meyer, E. & Meyer, K. F. (1952). Studies on immunisation against plague. I. The isolation and characterisation of the soluable antigen of Pasteurella pestis. Journal of Immunology 68, 131–145.CrossRefGoogle Scholar
Barnes, A. M. & Poland, J. D. (1983). Plague in the United States. 1982. Morbidity and Mortality Weekly Report, Centres for Disease Control Surveillance Summaries 32, 1988.Google ScholarPubMed
Bennett, L. G. & Tornabene, T. G. (1974). Characterisation of the antigenic subunits of the envelope protein of Yersinia pestis. Journal of Bacteriology 117, 48–55.CrossRefGoogle ScholarPubMed
Bolin, I., Portnoy, D. A. & Wolf-Watz, H. (1985). Expression of the temperature-inducible outer membrane proteins of Yersiniae. Infection and Immunity 48, 234–240.CrossRefGoogle ScholarPubMed
Carniel, E., Mazigh, D. & Mollaret, H. H. (1987). Expression of iron-related proteins in Yersinia species and their relation to virulence. Infection and Immunity 55, 277–280.CrossRefGoogle Scholar
Carter, P. B., Zahorchak, R. J. & Brubaker, R. R. (1980). Plague virulence antigens from Yersinia enterocolitica. Infection and Immunity 28, 638–640.CrossRefGoogle ScholarPubMed
Fox, E. N. & Higuchi, K. (1958). Synthesis of the Fraction 1 antigenic protein by Pasteurella pestis. Journal of Bacteriology 75, 209–216.CrossRefGoogle Scholar
Hudson, B. W., Kartman, L. & Prince, F. M. (1966). Pasteurella pestis detection in fleas by fluorescent antibody staining. Bulletin of the World Health Organisation 34, 709–714.Google ScholarPubMed
Hudson, B. W., Goldenberg, M. I., McCluskie, J. D., Larson, H. E., McGuire, C. D., Barnes, A. M. & Poland, J. D. (1971). Serological and bacteriological investigations of an outbreak of plague in an urban tree squirrel population. American Journal of Tropical Medicine and Hygiene 20, 255–263.CrossRefGoogle Scholar
Barnes, A. M. & Poland, J. D. (1971). Serological and bacteriological investigations of an outbreak of plague in an urban tree squirrel population. American Journal of Tropical Medicine and Hygiene 20, 255–263.Google Scholar
Kartman, L. (1960). The role of rabbits in sylvatic plague epidemiology, with special attention to human cases in New Mexico and use of the fluorescent antibody technique for detection of Pasteurella pestis in field specimens. Zoonoses Research 1, 1–27.Google Scholar
Mazza, G., Karu, A. E. & Kingsbury, D. T. (1985). Immune response to plasmid- and chromosome- encoded Yersinia antigens. Infection and Immunity 48, 676–685.CrossRefGoogle ScholarPubMed
Moody, M. D. & Winter, C. C. (1959). Rapid identification of Pasteurella pestis with fluorescent antibody. III. Staining Pasteurella pestis in tissue impression smears. Journal of Infectious Diseases 104, 288–294.CrossRefGoogle ScholarPubMed
Nairn, R. C. (1976). Fluorescent Protein Tracing, 4th edn, p. 163. Edinburgh: Churchill Livingstone.Google Scholar
Phillips, A. P. (1984). An alternative to PPD for reducing the bleaching of fluorescein, evaluated in bacterial immunofluorescence assays. Journal of Immunological Methods 66, 189–190.CrossRefGoogle ScholarPubMed
Phillips, A. P. & Martin, K. L. (1982 a). Evaluation of a microfluorometer in immunofluorescence assays of individual spores of Bacillus anthracis and Bacillus cereus. Journal of Immunological Methods 49, 271–282.CrossRefGoogle ScholarPubMed
Phillips, A. P. & Martin, K. L. (1982 b). Assessment of immunofluorescence measurements of individual bacteria in direct and indirect assays for Bacillus anthracis and Bacillus cereus spores. Journal of Applied Bacteriology 53, 223–231.CrossRefGoogle ScholarPubMed
Phillips, A. P. & Martin, K. L. (1985). Dual parameter scatter-flow analysis of Bacillus spores. Cytometry 6, 124–129.CrossRefGoogle ScholarPubMed
Phillips, A. P., Martin, K. L. & Capey, A. J. (1987). Direct and indirect immunofluorescence analysis of bacterial populations by flow cytometry. Journal of Immunological Methods 101, 219–228.CrossRefGoogle ScholarPubMed
Quan, S. F., Knapp, W., Goldenberg, M. I., Hudson, B. W., Lawton, W. D., Chen, T. H. & Kartman, L. (1965). Isolation of a strain of Pasteurella pseudotuberculosis from Alaska identified as Pasteurella pestis: an immunofluorescent false positive. American Journal of Tropical Medicine and Hygiene 14, 424–432.CrossRefGoogle ScholarPubMed
Shepherd, A. J., Hummitzsch, D. E., Leman, P. A., Swanepoel, R. & Searle, L. A. (1986). Comparative tests for detection of plague antigen and antibody in experimentally infected wild rodents. Journal of Clinical Microbiology 24, 1075–1078.CrossRefGoogle ScholarPubMed
Walker, R. V. (1962). Studies on the immune response of guinea pigs to the envelope substance of Pasteurella pestis. I. Immunogenicity and persistance of large doses of Fraction 1 in guinea pigs observed with fluorescent antibody. Journal of Immunology 88, 153–163.CrossRefGoogle Scholar
Williams, J. E. & Cavanaugh, D. C. (1983). Chronic infections in laboratory rodents from inoculation of non-encapsulated plague bacilli (Yersinia pestis). Experimentia 39, 408–409.CrossRefGoogle Scholar
Williams, J. E. & Cavanaugh, D. C. (1984). Potential for rat plague from non-encapsulated variants of the plague bacillus (Yersinia pestis). Experimentia 40, 739–740.CrossRefGoogle Scholar
Williams, J. E., Gentry, M. K., Braden, C. A., Leister, F. & Yolken, R. H. (1984). Use of an enzyme-linked immunosorbent assay to measure antigenaemia during acute plague. Bulletin of the World Health Organisation 62, 463–466.Google ScholarPubMed
Williams, J. E., Arntzen, L., Tyndal, G. L. & Isaacson, M. (1986). Application of enzyme immunoassays for the confirmation of clinically suspect plague in Namibia, 1982. Bulletin of the World Organisation 64, 745–752.Google ScholarPubMed
Winter, C. C. & Moody, M. D. (1959). Rapid identification of Pasteurella pestis with fluorescent antibody. II. Specific identification of Pasteurella pestis in dried smears. Journal of Infections Diseases 104, 281–287.CrossRefGoogle ScholarPubMed
Winter, C. C., Cherry, W. B. & Moody, M. D. (1960). An unusual strain of Pasteurella pestis isolated from a fatal human case of plague. Bulletin of the World Health Organisation 23, 408–409.Google ScholarPubMed