Development and evaluation of aptamer magnetic capture assay in conjunction with real-time PCR for detection of Campylobacter jejuni

作者：Suh Soo Hwan; Dwivedi Had P^*; Jaykus Lee Ann

来源：LEBENSMITTEL-WISSENSCHAFT UND-TECHNOLOGIE-FOOD SCIENCE AND TECHNOLOGY, 2014, 56(2): 256-260.

DOI：10.1016/j.lwt.2013.12.012

• Aptamer
• Aptamer magnetic capture
• Campylobacter jejuni
• Real-time PCR
• Pathogen detection

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.

• 出版日期2014-5