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American Society for Microbiology, Applied and Environmental Microbiology, 5(70), p. 3183-3187, 2004

DOI: 10.1128/aem.70.5.3183-3187.2004

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Multiplex PCR Assay for Detection of Bacterial Pathogens Associated with Warm-Water Streptococcosis in Fish

Journal article published in 2004 by Mata Ai, A. I. Mata, A. Gibello, A. Casamayor, M. M. Blanco, L. Domínguez, J. F. Fernández-Garayzábal ORCID
This paper is made freely available by the publisher.
This paper is made freely available by the publisher.

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Abstract

ABSTRACT 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 , Streptococcus parauberis , 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.