Characterization of ISR region and development of a PCR assay for rapid detection of the fish pathogen Tenacibaculum soleae
| Autor: | J.I. Navas, Roberto de la Herrán, Abdel Mounim Hamman-Khalifa, Jose R. López |
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| Rok vydání: | 2011 |
| Předmět: |
DNA
Bacterial food.ingredient Molecular Sequence Data Biology Polymerase Chain Reaction Microbiology law.invention Fish Diseases chemistry.chemical_compound food Flavobacteriaceae Infections law RNA Ribosomal 16S DNA Ribosomal Spacer Genetics Animals Agar Molecular Biology Gene Pathogen Polymerase chain reaction Detection limit Base Sequence Fishes 16S ribosomal RNA biology.organism_classification Molecular biology Bacterial Typing Techniques Tenacibaculum RNA Ribosomal 23S chemistry DNA Bacteria |
| Zdroj: | FEMS Microbiology Letters. 324:181-188 |
| ISSN: | 0378-1097 |
| DOI: | 10.1111/j.1574-6968.2011.02404.x |
| Popis: | The aims of this work were to characterize the 16S-23S internal spacer region of the fish pathogen Tenacibaculum 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 16S 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. |
| Databáze: | OpenAIRE |
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