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PCR‐ELISA detection of Escherichia coli in milk
Author(s) -
Daly P.,
Collier T.,
Doyle S.
Publication year - 2002
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1046/j.1472-765x.2002.01074.x
Subject(s) - escherichia coli , pasteurization , amplicon , biology , mastitis , microbiology and biotechnology , polymerase chain reaction , contamination , enterobacteriaceae , dna extraction , food science , gene , biochemistry , ecology
Aims: The purpose of this study was to develop a reliable molecular procedure for the detection of Escherichia coli in milk. Methods and Results: Robust and expeditious DNA extraction and PCR techniques were evaluated using Enzyme‐Linked Immunosorbent Assay (ELISA) detection of biotin‐labelled amplicons to facilitate optimal detection of E. coli DNA. Conclusions: It was found that 5 E. coli colony‐forming units (cfu) could be detected per PCR reaction using the PCR‐ELISA system, equating to a sensitivity of detection of 100 E. coli cfu ml −1 pasteurized milk. Significance and Impact of the Study: This approach should facilitate evaluation of milk contamination and enable rapid detection of E. coli mastitis, leading to correct deployment of relevant antibiotic therapy and improved animal welfare.