Open AccessCharacterization of ISR region and development of a PCR assay for rapid detection of the fish pathogen T enacibaculum soleae
Author(s) -
López Jose R.,
HammanKhalifa Abdel M.,
Navas José I.,
la Herran Roberto
Publication year - 2011
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2011.02404.x
Subject(s) - pathogen , biology , internal transcribed spacer , 16s ribosomal rna , polymerase chain reaction , 23s ribosomal rna , microbiology and biotechnology , ribosomal rna , gene , fish <actinopterygii> , intraspecific competition , dna , bacteria , genetics , zoology , rna , fishery , ribosome
The aims of this work were to characterize the 16 S –23 S internal spacer region of the fish pathogen T enacibaculum soleae and to develop a PCR assay for its identification and detection. All T . soleae strains tested displayed a single internal spacer region class, containing tRNA I le and tRNA A la genes; nevertheless, a considerable intraspecific heterogeneity was observed. However, this region proved to be useful for differentiation of T . soleae from related and non‐related species. Species‐specific primers were designed targeting the 16 S rRNA gene and the internal spacer region region, yielding a 1555‐bp fragment. Detection limit was of 1 pg DNA per reaction (< 30 bacterial cells) when using pure cultures. The detection level in the presence of DNA from fish or other bacteria was lower; however, 10 pg were detected at a target/background ratio of 1 : 10 5 . The PCR assay proved to be more sensitive than agar cultivation for the detection of T . soleae from naturally diseased fish, offering a useful tool for diagnosis and for understanding the epidemiology of this pathogen.