A specific DNA hybridization probe for detection of Pasteurella piscicida

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Abstract
A 692 base pair (bp) DNA fragment from the bacterial fish pathogen Pasteurella piscicida was cloned into pUC9 and used as a hybridization probe for the identification of P. piscicida. The radiolabeled probe hybridized with itself, but did not hybridize in colony hybridization with other fish pathogens (including Aeromonas hydrophila, A. salmonicida, Edwardsiella tarda, Pseudomonas anguilliseptica, Vibrio anguillarum and Yersinia ruckeri), or with other related organisms (Haemophilus influenzae, Pasteurella haemolytica, and P. multocida). Photobiotin-labeled probe hybridization detected P. piscicida chromosomal DNA by Southern and dot blot hybridizations. The radiolabeled probe directly detected P. piscicida on nitrocellulose filters smeared with infected kidney and spleen tissue of yellowtail Seriola quinqueradiata. The 32P-labeled DNA probe was capable of detecting 3.9 ng of purified DNA and 105 cells of P. piscicida. The cloned specific DNA probe can be used for rapid detection and identification of P. piscicida.

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