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Evaluation of PCR primers targeting the gro EL gene for the specific detection of Streptococcus agalactiae in the context of aquaculture
Author(s) -
Leigh W.J.,
Zadoks R.N.,
Jaglarz A.,
Costa J.Z.,
Foster G.,
Thompson K.D.
Publication year - 2018
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/jam.13925
Subject(s) - streptococcus iniae , streptococcus agalactiae , biology , microbiology and biotechnology , polymerase chain reaction , context (archaeology) , pathogen , aquaculture , bacteria , gene , genetics , streptococcus , fish <actinopterygii> , fishery , paleontology
Abstract Aims The aim of this study was to design a set of primers for specific detection and identification of Streptococcus agalactiae in polymerase chain reaction ( PCR ) that can detect a diverse range of S. agalactiae isolates from different hosts and that it is capable of discriminating between S. agalactiae and other species that are closely related or potentially present in aquaculture environments, notably Streptococcus iniae . Methods and Results Primers, based on the gro EL 2 gene of S. agalactiae, were shown to be epidemiologically sensitive to 97 isolates of S. agalactiae , representing 11 clonal complexes derived from piscine, terrestrial and aquatic mammalian host species. The primers were tested with 10 S. iniae isolates and 22 other comparator species with no cross‐reaction observed after optimization of reaction conditions. They have a high analytical sensitivity, detecting as few as 10 copies of S. agalactiae genomic DNA per reaction and are capable of detecting the target in DNA extracted from the brains of infected fish. Conclusions The primers proved suitable for the sensitive and specific detection of S. agalactiae from dairy‐, human‐ and fish‐related origins by PCR . Significance and Impact of the Study Due to the importance of S. agalactiae as a pathogen, many PCR primers have been published for this bacterium, designed largely for its detection in dairy and human samples, but many cross‐reacting with S. iniae . The ability to differentiate between S. agalactiae and S. iniae in aquaculture derived samples is important as both infect fish, causing similar disease symptoms and are phenotypically similar, yet control strategies and zoonotic risk are species specific.