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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

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

Full text
Author(s):
Rubio, Marcela da Silva [1] ; Casarin Penha Filho, Rafael Antonio [1] ; de Almeida, Adriana Maria [1] ; Berchieri Junior, Angelo [1]
Total Authors: 4
Affiliation:
[1] Sao Paulo State Univ FCAV UNESP, Sch Agr & Vet Sci, Sao Paulo - Brazil
Total Affiliations: 1
Document type: Journal article
Source: AVIAN PATHOLOGY; v. 46, n. 6, p. 644-651, 2017.
Web of Science Citations: 2
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 detect both host-specific biovars but not other serovars and pSG and pSP differentiate biovars. Three amplicons showed different melting temperatures (Tm), allowing differentiation. The pSGP amplicon (97 bp) showed Tm of 78 degrees C for both biovars. The pSG amplicon (273 bp) showed a Tm of 86.2 degrees C for S. Gallinarum and pSP amplicon (260 bp) dissociated at 84.8 degrees C for S. Pullorum identification. The multiplex qPCR in real time showed high sensitivity and was capable of quantifying 10(8)-10(1) CFU of these biovars. (AU)

FAPESP's process: 14/05496-7 - Use of quantitative Real-Time PCR for differential diagnosis of Salmonella enterica subsp. enterica serovar Enteritidis, Typhimurium and Gallinarum (biovars Gallinarum and Pullorum) in domestic poultry (Gallus Gallus)
Grantee:Marcela da Silva Rubio
Support Opportunities: Scholarships in Brazil - Doctorate
FAPESP's process: 12/24017-7 - Evaluation of the infection and immunity in susceptible and resistant lines of chicken (Gallus gallus) challenged with Salmonella enterica subsp. enterica sorovar Gallinarum
Grantee:Rafael Antonio Casarin Penha Filho
Support Opportunities: Scholarships in Brazil - Post-Doctoral