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Original Article

J App Pharm Sci. 2018; 8(3): 126-132

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.


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.

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

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