Journal
LWT-FOOD SCIENCE AND TECHNOLOGY
Volume 56, Issue 2, Pages 256-260Publisher
ELSEVIER
DOI: 10.1016/j.lwt.2013.12.012
Keywords
Aptamer; Aptamer magnetic capture; Campylobacter jejuni; Real-time PCR; Pathogen detection
Categories
Funding
- Food Microbiology Committee of the North American Branch of the International Life Sciences Institute (ILSI)
- ILSI North America
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A prototype method for the concentration and detection of Campylobacter jejuni was developed using a previously reported biotinylated DNA aptamer in conjunction with qPCR. The so-called aptamer-based magnetic capture-qPCR (AMC-qPCR) assay was compared to a similar immuno-magnetic separation (IMS)-qPCR assay. In small volume experiments (300 mu l) applied to serially diluted C. jejuni suspended in buffer containing a mixed culture of other common food borne pathogens, the lower detection limit of the AMC-qPCR method was 1.1 log(10)/300 mu l C. jejuni cells, one log(10) better (lower) than that of IMS-qPCR (2.1 log(10) CFU/300 mu l). AMC-qPCR capture efficiency was 10-13% at assay detection limit. In 10 ml scale-up experiments, the lower detection limit of AMC-qPCR was 2.0 log(10) CFU/10 ml with corresponding capture efficiency of 4-7%. Nucleic acid aptamers are promising alternatives to antibodies for magnetic bead-based capture followed by qPCR detection. (C) 2013 Elsevier Ltd. All rights reserved.
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