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The food items processor vessel couldn't be warmth-sterilized, so right after washing with soap and cleansing with sodium hypochlorite and ethanol, it absolutely was left inside the UV cupboard (λ = 254 nm) for five min.
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This get the job done aimed to investigate the microbiological high-quality of ninety five samples of Cotija cheese and sixteen samples of Bola de Ocosingo cheese from the qualified detection of 5 pathogenic microorganisms of fascination in dairy foods: Salmonella enterica, Escherichia coli enterotoxigenic (ETEC), E.
five mL with the intermediate period following the centrifugation with the homogenized cheese on peptone h2o as explained in Portion two.six. Cultures ended up incubated beneath the respective problems for each pathogenic bacterium (Table one). Afterward, each society was warmth-inactivated, and DNA extraction and qPCR have been carried out in duplicate following the course of action explained earlier mentioned. An additional set of experiments was performed following the same methodology but spiking a warmth-inactivated inoculum.
The amplification of DNA from the pure cultures of strains produced amplification curves with threshold cycle (Ct) values between eleven and fourteen. When DNA from microorganisms aside from the concentrate on was utilised as being a template, no amplification or curves deposit pulsa tanpa potongan with Ct values better than 30 have been acquired, that may be attributed for the development of primer dimers or other non-specific solutions [34].
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The gene sequences for toxin and antigenic proteins precise to each pathogenic bacterium have been thought of targets in the design on the primers and probes Employed in qPCR (Table 2).