Detection of Campylobacter DNA using magnetic nanoparticles coupled with PCR and a colorimetric end-point system

Abstract

Campylobacter is an important food-borne pathogen causing acute gastroenteritis worldwide. Magnetic nanoparticle-based PCR coupled with streptavidin-horseradish peroxidase and a substrate was used for colorimetric detection. Forward primers conjugated to magnetic nanoparticles facilitated separation and concentration of Campylobacter DNA in a sample matrix. After PCR, a green color developed and was observed using the unaided eye, or detected using a spectrophotometer. High specificity and sensitivity of the 100 fg DNA/PCR reaction were achieved in pure culture experiments. The technique was applied for detection of Campylobacter on naturally contaminated chicken skin. All positive results were in agreement with results achieved using a conventional culture method. The magnetic nanoparticle-PCR-enzyme linked gene assay was practical and useful for detection of Campylobacter in complex matrices with PCR-interfering substances. © 2016, The Korean Society of Food Science and Technology and Springer Science+Business Media Dordrecht.