A multiplex polymerase chain reaction protocol for the detection of Photobacterium damselae and subspecies piscicida and damselae discrimination, with internal amplification control, was developed. Assay specificity was assessed by testing 19 target and 25 non-target pure cultures. The detection limit was 500 fg, corresponding to 100 genome equivalents. The optimized protocol was also prevalidated with spleen, kidney and blood samples from infected and uninfected sea bass, without any culture step, and it can be proposed as a valid alternative to culture standard methods for the rapid and specific diagnosis of photobacteriosis in fish. © 2009 Blackwell Publishing Ltd.
Development of a multiplex PCR assay for photobacterium damselae subsp. piscicida identification in fish samples / Amagliani, G.; Omiccioli, E.; Andreoni, F.; Boiani, R.; Bianconi, I.; Zaccone, R.; Mancuso, M.; Magnani, M.. - In: JOURNAL OF FISH DISEASES. - ISSN 0140-7775. - ELETTRONICO. - 32:8(2009), pp. 645-653. [10.1111/j.1365-2761.2009.01027.x]
Development of a multiplex PCR assay for photobacterium damselae subsp. piscicida identification in fish samples
Bianconi I.;
2009-01-01
Abstract
A multiplex polymerase chain reaction protocol for the detection of Photobacterium damselae and subspecies piscicida and damselae discrimination, with internal amplification control, was developed. Assay specificity was assessed by testing 19 target and 25 non-target pure cultures. The detection limit was 500 fg, corresponding to 100 genome equivalents. The optimized protocol was also prevalidated with spleen, kidney and blood samples from infected and uninfected sea bass, without any culture step, and it can be proposed as a valid alternative to culture standard methods for the rapid and specific diagnosis of photobacteriosis in fish. © 2009 Blackwell Publishing Ltd.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione
