Endpoint and real-time PCR optimization for detection of Salmonella enterica serovar Gallinarum in poultry from Costa Rica
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molecular detection, DNA, taxonomic placement, Enterobacteriaceae

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Leza-LezaM. T., Víquez-RuizE., Barquero-CalvoE., Sancho-BlancoC., & Umaña-CastroR. (2022). Endpoint and real-time PCR optimization for detection of Salmonella enterica serovar Gallinarum in poultry from Costa Rica. UNED Research Journal, 14(1), e3831. https://doi.org/10.22458/urj.v14i1.3831


Introduction: Avian typhoid and pulorosis, diseases caused by Salmonella enterica subsp. enterica serotype Gallinarum biotype Gallinarum and biotype Pullorum, respectively, are responsible for high mortality in poultry, generating significant economic losses for poultry farmers. Objective: To strengthen the detection of the causative agents of Fowl typhoid and pullorum disease by optimizing molecular techniques, such as endpoint PCR and real-time PCR (qPCR). Methods: Bacterial strains control groups, isolates, and tissue from poultry infected with Salmonella Gallinarum were used to standardize the detection by endpoint PCR and qPCR. Results: For the endpoint PCR, a repeatability, specificity, and sensitivity of 100% was obtained, and a Kappa value of 0.98 for reproducibility. With the qPCR, an efficiency of 103% was obtained, while the coefficients of variation for repeatability and reproducibility were lower than 6%. The detection limit of genomic DNA was 6.4 pg/μL, and the number of viable cells was 3x102 CFU/mL for endpoint PCR and 10 copies of DNA per reaction for qPCR. Through sequencing and analysis of the molecular taxonomy of the PCR products, the strains in this study were positioned together with those of serotype S. Gallinarum, corroborating their identity. Conclusion: It was possible to optimize a molecular technique that allows a rapid, reliable, and sensitive detection of Salmonella Gallinarum /Pullorum, which can reduce the waiting time to take action in cases of clinical suspicion and possible outbreaks.
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