PCR-ELISA detection of Escherichia coli in milk.
OBJECTIVE
The purpose of this study was to develop a reliable molecular procedure for the detection of Escherichia coli in milk.


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.


CONCLUSIONS
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.
Daly
P
P
Department of Biology, National University of Ireland, Maynooth, Kildare, Ireland.
Collier
T
T
Doyle
S
S
D000818:Animals; D002417:Cattle; D004587:Electrophoresis, Agar Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification*; D005838:Genotype; D008892:Milk / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*