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

doi:10.1128/JCM.39.7.2558-2564.2001

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.

Tables

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Table 1.
Comparison of sorbitol fermentation results obtained using different test media
Sorbitol fermentation result No. of isolates as determined by:
APF-20E PRAS Andrades
Positive 18 10 16
Negative 14 12 4
Weak 0 13 11^a
Discrepant reaction on repeat testing 3 ND^b 7^a
↵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 result^a No. of isolates in PCR fingerprint group: No. of isolates with α-Glu activity:
I II III Positive Negative
Negative 13 0 0 13 0
Positive 8 4 7 10 9
Uncertain 3 0 0 3 0
↵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 activity No. of isolates in PCR fingerprint group:
I II III
Positive 24 0 2
Negative 0 4 5