Research Article | Volume: 8, Issue: 3, March, 2018

Chromocult Coliform agar and duplex PCR assays as methodologies for tracking Escherichia coli K12 in industrial biotechnological processes

Gisele d’Angelis Antunes,  Janine Aparecida Correia Durães Gandra,  Elane Alquimim Moreira,  Wesley Cézar Silva Machado,  Shirley da Silva Gomes Magalhães,  Mauro Aparecido de Sousa Xavier,  Alessandra Rejane Ericsson de Oliveira Xavier   
Gisele d’Angelis Antunes1, Janine Aparecida Correia Durães Gandra2, Elane Alquimim Moreira1, Wesley Cézar Silva Machado1, Shirley da Silva Gomes Magalhães1, Mauro Aparecido de Sousa Xavier2, Alessandra Rejane Ericsson de Oliveira Xavier2

1Novo Nordisk Produção Farmacêutica do Brasil, ALP Production, Montes Claros, MG, Brazil.

2State University of Montes Claros, Center for Biological and Health Sciences, Graduate Program in Biotechnology, Microbiology Laboratory, Montes Claros, MG, Brazil.

Open Access   

Published:  Mar 30, 2018

DOI: 10.7324/JAPS.2018.8318
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Abstract

The objective of this work was to establish support for the validation of the microbiological method associated with the duplex PCR assay for Escherichia coli K12 in waste and intermediate products of an industrial biotechnological process. For this purpose, samples of Escherichia coli K12 from critical control points known as Killing System (KS) and Germ Filtration (GF) were analyzed. Experiments to determine the microbial load of E. coli K12 based on the direct plating and filter membrane methods revealed the absence of this microorganism in the KS and GF samples. The chromogenic medium was able to recover strains of E. coli inoculated intentionally in GF samples, but the same result was not possible in samples from KS. The robustness of the incubation time of the KS and GF samples were calculated by the ANOVA test and the results showed no statistically significant difference between them (p-value 0.181 and 0.733). Duplex PCR analysis was shown to be able to differentiate a standard strain of Escherichia coli ATCC 8739 from Escherichia coli K12. The results will be used as a guideline for validation of a method for E. coli K12 tracking in an industrial biotechnological process.


Keyword:     Chromocult® Coliform Agar Filter Membrane Escherichia coli K12 duplex PCR Genetically Modified Organism Detection.

Citation:

Antunes GA, Gandra JACD, Moreira EA, Machado WCS, Magalhães SSG, Xavier MAS, Xavier AREO. Chromocult Coliform agar and duplex PCR assays as methodologies for tracking Escherichia coli K12 in industrial biotechnological processes. J App Pharm Sci, 2018; 8(03): 126-132.

Copyright: © The Author(s). This is an open-access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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