Journal
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
Volume 70, Issue 5, Pages 3183-3187Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/AEM.70.5.3183-3187.2004
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A multiplex PCR-based method was designed for the simultaneous detection of the main pathogens involved in Warm-water streptococcosis in fish (Streptococcus iniae, Streptococcus difficilis, Sireptococcus paratiberis, and Lactococcus garvieae). Each of the four pairs of oligonucleotide primers exclusively amplified the targeted gene of the specific microorganism. The sensitivity of the multiplex PCR using purified DNA was 25 pg for S. iniae, 12.5 pg for S. difficilis, 50 pg for S. parauberis, and 30 pg for L. garvieae. The multiplex PCR assay was useful for the specific detection of the four species of bacteria not only in pure culture but also in inoculated fish tissue homogenates and naturally infected fish. Therefore, this method could be a useful alternative to the culture-based method for the routine diagnosis of warm-water streptococcal infections in fish.
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