Development of a multiplex qPCR in real time for quantification and differential diagnosis of Salmonella Gallinarum and Salmonella Pullorum.

Development of a multiplex qPCR in real time for quantification and differential diagnosis of Salmonella Gallinarum and Salmonella Pullorum. Avian Pathol. 2017 Jun 07;:1-31 Authors: Rubio MDS, Penha Filho RAC, Almeida AM, Berchieri Junior A Abstract Currently there are 2659 Salmonella serovars. The host-specific biovars Salmonella Pullorum and Salmonella Gallinarum, cause systemic infections in food-producing and wild birds. Fast diagnosis is crucial to control the dissemination in avian environments. The present work describes the development of a multiplex qPCR in real time using a low cost DNA dye (SYBr Green) to identify and quantify these biovars. Primers were chosen based on genomic regions of difference (RoD) and optimized to control dimers. Primers pSGP differentiate from other serovars and pSG and pSP differentiate biovars. Three amplicons showed different melting temperatures (Tm) allowing differentiation. pSGP amplicon (97bp) showed Tm of 78°C for both biovars. pSG amplicon (273bp) showed a Tm of 86.2°C for S. Gallinarum and pSP amplicon (260bp) dissociated at 84.8°C for S. Pullorum identification. The multiplex qPCR in real time showed high sensitivity and was capable to quantify 10(8) to 10(1) CFU of these biovars. PMID: 28589774 [PubMed - as supplied by publisher]
Source: Avian Pathology - Category: Pathology Authors: Tags: Avian Pathol Source Type: research