Development of a PCR assay for Streptococcus iniae based on the lactate oxidase (lctO) gene with potential diagnostic value

Decreasing levels of the fish pathogen Streptococcus iniae following inoculation into the sludge digester of a zero-discharge recirculating aquaculture system (RAS)

“…The 870-bp band was excised and sequenced by a commercial laboratory (Hylabs, Rehovot, Israel). The sequence was identical to the expected S. iniae gene for lactate oxidase (Mata et al, 2004b; data not shown).…”

“…PCR amplification was performed in a 50-µL reaction volume containing DNA template (80 ng) using the LOX 1/2 primer set, which specifically amplifies the gene encoding lactate oxidase, producing an 870-bp (Mata et al, 2004b) or 920-bp (Nawawi et al, 2009) …”

“…Several PCR-based assays have been developed for the detection of S. iniae (Berridge et al, 1998;Lau et al, 2004;Mata et al, 2004a,b;Zlotkin et al, 1998). In the present study, we applied the method described by Mata et al (2004b), based on amplification of the gene encoding lactate oxidase, to examine the survival of S. iniae in the sludge of a UASB reactor in a pilot-scale RAS system, as well as its presence in sludge effluent from a commercial RAS.…”

Induction and characterization of lysogenic bacteriophages fromStreptococcus iniae

“…iniae using the primer combination LOX‐1/LOX‐2 (Mata et al . ). Streptococcus iniae isolates were preserved at −80°C using the Microbank porous bead system, and propagated aerobically at 28°C, with continuous agitation if in broth culture.…”

Streptococcus iniae outbreaks in Brazilian Nile tilapia (Oreochromis niloticus L: ) farms

“…The colonies were collected, diluted in 180 μL of lysis solution (20 mg ml -1 lisozyme; 20 mM Tris-HCl, pH 8.0; 2 mM EDTA and 1.2% Triton ® ), and incubated at 37 °C for 1 h. Total bacterial DNA was then extracted using the commercial kit QIAamp DNA mini kit (Qiagen) according to the manufacturer's instructions. To confirm the diagnosis, extracted DNA was used as a template in a S. iniae- specific PCR performed according to the procedure previously described by Mata et al (12). In addition, 16S rRNA of three randomly selected strains (SI25-06, SI27-06 and SI29-06) were amplified by PCR with the universal primers C70 and B37 according to the method described by Fox et al (10).…”