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Journal of Clinical Microbiology
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Bacteriology

Pasteurellamultocida subsp.multocida and P. multocida subsp.septica Differentiation by PCR Fingerprinting and α-Glucosidase Activity

Sharon Hunt Gerardo, Diane M. Citron, Marina C. Claros, Helen T. Fernandez, Ellie J. C. Goldstein
Sharon Hunt Gerardo
R. M. Alden Research Laboratory, Santa Monica/UCLA Medical Center, Santa Monica, California 90404;
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Diane M. Citron
R. M. Alden Research Laboratory, Santa Monica/UCLA Medical Center, Santa Monica, California 90404;
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Marina C. Claros
Institute of Medical Microbiology, University of Leipzig, Leipzig, Germany; and
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Helen T. Fernandez
R. M. Alden Research Laboratory, Santa Monica/UCLA Medical Center, Santa Monica, California 90404;
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Ellie J. C. Goldstein
R. M. Alden Research Laboratory, Santa Monica/UCLA Medical Center, Santa Monica, California 90404;
University of California School of Medicine, Los Angeles, California 90095
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DOI: 10.1128/JCM.39.7.2558-2564.2001
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    Fig. 1.

    P. multocida subsp.septica PCR fingerprint profiles. (A) Thirteen sorbitol-negative, α-Glu-positive and three sorbitol-uncertain, α-Glu-positive P. multocida bite wound isolates, examined by PCR fingerprint analyses using the single primer, M13 core, exhibited the group I PCR fingerprint profile characteristic ofP. multocida subsp. septica. (Sorbitol fermentation was based on a consensus of API-20E, PRAS, and Andrades sorbitol fermentation reactions.) Lane 1, negative control (no DNA template); lane 2, DNA ladder; lane 3, P. multocidasubsp. multocida ATCC 12947; lane 4, P. multocida subsp. septica ATCC 51688; lanes 5 to 20, bite wound isolates. (B) Seven sorbitol-positive, α-Glu-positiveP. multocida bite wound isolates, examined by PCR fingerprint analyses using the single primer, M13 core, showed the group I PCR fingerprint profile characteristic of the P. multocida subsp. septica control strain. Lane 1, negative control (no DNA template); lanes 2 and 10, DNA ladder; lanes 3 to 9, bite wound isolates; lane 11, P. multocida subsp.septica ATCC 51688.

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    Fig. 2.

    P.multocida subsp.multocida PCR fingerprint analyses. (A) Nine sorbitol-positive, α-Glu-negative P. multocida subsp.multocida bite wound isolates, examined by PCR fingerprint analyses using the single primer, M13 core, displayed either the group II or group III PCR fingerprint profile. (Sorbitol fermentation was based on a consensus of API-20E, PRAS, and Andrades sorbitol fermentation reactions.) Lane 1, DNA ladder; lane 2, P. multocida subsp. septica ATCC 51688; lane 3,P. multocida subsp. multocida ATCC 12947; lanes 4 to 7, bite wound isolates demonstrating the group II PCR fingerprint pattern; lanes 8 to 12, bite wound isolates showing the group III PCR fingerprint profile; lane 13, negative control (no DNA template). (B) Two sorbitol-positive, α-Glu-positive P. multocida bite wound isolates, examined by PCR fingerprinting analyses using the single primer, M13 core, exhibited the group III PCR fingerprint profile characteristic of other P. multocidasubsp. multocida clinical isolates. Lane 1, negative control (no DNA template); lane 2, DNA ladder; lane 3, P. multocida subsp. septica ATCC 51688; lane 4,P. multocida subsp. multocida ATCC 12947; lanes 5 and 6, bite wound isolates.

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  • Table 1.

    Comparison of sorbitol fermentation results obtained using different test media

    Sorbitol fermentation resultNo. of isolates as determined by:
    APF-20EPRASAndrades
    Positive181016
    Negative14124
    Weak01311a
    Discrepant reaction on repeat testing3NDb7a
    • ↵a Overlapping groups.

    • ↵b ND, not done.

  • Table 2.

    Sorbitol fermentation reactions show poor correlation with PCR fingerprint analyses and α-Glu activity

    Sorbitol fermentation resultaNo. of isolates in PCR fingerprint group:No. of isolates with α-Glu activity:
    IIIIIIPositiveNegative
    Negative1300130
    Positive847109
    Uncertain30030
    • ↵a Based on consensus of API-20E, PRAS, and Andrades sorbitol fermentation reactions.

  • Table 3.

    α-Glu activity shows good correlation with the PCR fingerprint profiles

    α-Glucosidase activityNo. of isolates in PCR fingerprint group:
    IIIIII
    Positive2402
    Negative045
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Pasteurellamultocida subsp.multocida and P. multocida subsp.septica Differentiation by PCR Fingerprinting and α-Glucosidase Activity
Sharon Hunt Gerardo, Diane M. Citron, Marina C. Claros, Helen T. Fernandez, Ellie J. C. Goldstein
Journal of Clinical Microbiology Jul 2001, 39 (7) 2558-2564; DOI: 10.1128/JCM.39.7.2558-2564.2001

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Pasteurellamultocida subsp.multocida and P. multocida subsp.septica Differentiation by PCR Fingerprinting and α-Glucosidase Activity
Sharon Hunt Gerardo, Diane M. Citron, Marina C. Claros, Helen T. Fernandez, Ellie J. C. Goldstein
Journal of Clinical Microbiology Jul 2001, 39 (7) 2558-2564; DOI: 10.1128/JCM.39.7.2558-2564.2001
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KEYWORDS

Bites and Stings
Pasteurella Infections
Pasteurella multocida
polymerase chain reaction
Wound Infection
alpha-Glucosidases

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