Short communication: Detection of stx2 and elt genes in bovine milk by using a multiplex PCR system.

Autor:	Nandi RDS; Department of Microbiology, Center for Biological Sciences, Universidade Estadual de Londrina, Londrina CEP 86051-980, Brazil., Campos AC; Department of Microbiology, Center for Biological Sciences, Universidade Estadual de Londrina, Londrina CEP 86051-980, Brazil., Puño-Sarmiento JJ; Department of Microbiology, Center for Biological Sciences, Universidade Estadual de Londrina, Londrina CEP 86051-980, Brazil., Maluta RP; Department of Microbiology, Center for Biological Sciences, Universidade Estadual de Londrina, Londrina CEP 86051-980, Brazil., Rocha SPD; Department of Microbiology, Center for Biological Sciences, Universidade Estadual de Londrina, Londrina CEP 86051-980, Brazil., Kobayashi RKT; Department of Microbiology, Center for Biological Sciences, Universidade Estadual de Londrina, Londrina CEP 86051-980, Brazil., Nakazato G; Department of Microbiology, Center for Biological Sciences, Universidade Estadual de Londrina, Londrina CEP 86051-980, Brazil. Electronic address: gnakazato@uel.br.
Jazyk:	angličtina
Zdroj:	Journal of dairy science [J Dairy Sci] 2017 Oct; Vol. 100 (10), pp. 7897-7900. Date of Electronic Publication: 2017 Aug 10.
DOI:	10.3168/jds.2017-13220
Abstrakt:	The aim of this study was to detect 2 important toxin genes from diarrheagenic Escherichia coli (DEC) in bovine milk using a new multiplex PCR. To standardize the multiplex PCR, the stx2 and elt genes were investigated for the detection of Shiga toxin-producing Escherichia coli (STEC) and enterotoxigenic E. coli (ETEC), respectively. The DNA template was prepared with a thermal procedure (boiling) and a commercial kit. Samples consisted of UHT and pasteurized milk, both skimmed, and STEC and ETEC were tested in concentrations between 10 1 and 10 9 cfu/mL. With the thermal procedure, the multiplex PCR system detected both pathotypes of E. coli at 10 9 cfu/mL in UHT and pasteurized milk. When the commercial kit was used for template preparation, STEC and ETEC could be detected at concentrations as low as 10 4 cfu/mL in UHT and pasteurized milk. Negative controls (Listeria monocytogenes, Salmonella Typhimurium, Salmonella Enteritidis, and Escherichia coli strain APEC 13) were not amplified with the multiplex PCR. These results indicate that the multiplex PCR was a rapid (less than 6 h) and efficient method to detect STEC and ETEC in milk using different methods for DNA preparation; however, the commercial kit was more sensitive than the thermal procedure. (Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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