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A serotype-specific polymerase chain reaction for identification of Pasteurella multocida serotype 1

Avian Diseases

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DOI: 10.1637/0005-2086(2002)046[0370:ASSPCR]2.0.CO;2

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Abstract

A serotype-specific polymerase chain reaction (PCR) assay was developed for detection and identification of Pasteurella multocida serotype 1, the causative agent of avian cholera in wild waterfowl. Arbitrarily primed PCR was used to detect DNA fragments that distinguish serotype 1 from the other 15 serotypes of P. multocida (with the exception of serotype 14). Oligonucleotide primers were constructed from these sequences, and a PCR assay was optimized and evaluated. PCR reactions consistently resulted in amplification products with reference strains 1 and 14 and all other serotype 1 strains tested, with cell numbers as low as 2.3 cells/ml. No amplification products were produced with other P. multocida serotypes or any other bacterial species tested. To compare the sensitivity and further test the specificity of this PCR assay with traditional culturing and serotyping techniques, tissue samples from 84 Pekin ducks inoculated with field strains of P. multocida and 54 wild lesser snow geese collected during an avian cholera outbreak were provided by other investigators working on avian cholera. PCR was as sensitive (58/64) as routine isolation (52/64) in detecting and identifying P. multocida serotype 1 from the livers of inoculated Pekins that became sick or died from avian cholera. No product was amplified from tissues of 20 other Pekin ducks that received serotypes other than type 1 (serotype 3, 12 × 3, or 10) or 12 control birds. Of the 54 snow geese necropsied and tested for P. multocida, our PCR detected and identified the bacteria from 44 compared with 45 by direct isolation. The serotype-specific PCR we developed was much faster and less labor intensive than traditional culturing and serotyping procedures and could result in diagnosis of serotype 1 pasteurellosis within 24 hr of specimen submission.

Additional Publication Details

Publication type:
Article
Publication Subtype:
Journal Article
Title:
A serotype-specific polymerase chain reaction for identification of Pasteurella multocida serotype 1
Series title:
Avian Diseases
DOI:
10.1637/0005-2086(2002)046[0370:ASSPCR]2.0.CO;2
Volume
46
Issue:
2
Year Published:
2002
Language:
English
Publisher:
American Association of Avian Pathologists
Publisher location:
Jacksonville, FL
Contributing office(s):
National Wildlife Health Center
Description:
8 p.
Larger Work Type:
Article
Larger Work Subtype:
Journal Article
Larger Work Title:
Avian Diseases
First page:
370
Last page:
377
Number of Pages:
8