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Development of a multiplex PCR assay for Photobacterium damselae subsp. piscicida identification in fish samples
Author(s) -
Amagliani G,
Omiccioli E,
Andreoni F,
Boiani R,
Bianconi I,
Zaccone R,
Mancuso M,
Magnani M
Publication year - 2009
Publication title -
journal of fish diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.819
H-Index - 85
eISSN - 1365-2761
pISSN - 0140-7775
DOI - 10.1111/j.1365-2761.2009.01027.x
Subject(s) - biology , multiplex polymerase chain reaction , subspecies , multiplex , sea bass , photobacterium , polymerase chain reaction , fish <actinopterygii> , microbiology and biotechnology , marine fish , gene , fishery , bacteria , vibrio , genetics , zoology
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.