Detection of Salmonella Enterica in Egg Yolk by PCR on a Microfluidic Disc Device Using Immunomagnetic Beads

Sensors

Shunsuke Furutani¹  Izumi Kubo²  Mitsutoshi Kajiya²  Narumi Aramaki² 

[1] Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-8-31 Midorigaoka, Ikeda, Osaka 563-8577, Japan;Graduate School of Engineering, Soka University, 1-236 Tangi, Hachioji, Tokyo 192-8577, Japan;

关键词: immunomagnetic beads;    salmonella enterica;    pcr;    inva;    egg yolk;   

DOI  :  10.3390/s20041060

来源: DOAJ

【 摘 要 】

Salmonella enterica is a pathogenic bacterium that causes foodborne illness. One of the vehicle foods of S. enterica are chicken eggs. Efficient collection of the bacterium is necessary to detect it specifically. We developed a method to detect S. enterica by PCR on a microfluidic disc device using a fluorescent probe. Salmonella enterica cells were isolated in the microchambers on the device, followed by thermal lysis and PCR targeting with the invA gene, a gene specific to S. enterica, were observed by measurement of the fluorescent signal that resulted from gene amplification. However, the developed method was unable to discriminate viable cells from dead cells. Consequently, in this study, magnetic beads modified with anti-Salmonella antibody were utilized to detect viable Salmonella cells from egg yolk prior to PCR on the device. While using the antibody-modified beads, egg yolk components, which inhibit PCR, were removed. The collected cells were subsequently detected by PCR of the invA gene on a microfluidic disc device. This method enabled the detection of viable cells without the inhibition of PCR by any egg component. S. enterica was detected at 5.0×10⁴ cells mL⁻¹ or at a higher concentration of egg yolk within 6 h including the sampling time.

【 授权许可】

Unknown