Detection of Salmonella Enterica in Egg Yolk by PCR on a Microfluidic Disc Device Using Immunomagnetic Beads
Salmonella <i>enterica</i> is a pathogenic bacterium that causes foodborne illness. One of the vehicle foods of <i>S. enterica</i> are chicken eggs. Efficient collection of the bacterium is necessary to detect it specifically. We developed a method to detect <i>S. enter...
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MDPI AG
2020-02-01
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| Series: | Sensors |
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| Online Access: | https://www.mdpi.com/1424-8220/20/4/1060 |
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| author | Izumi Kubo Mitsutoshi Kajiya Narumi Aramaki Shunsuke Furutani |
| author_facet | Izumi Kubo Mitsutoshi Kajiya Narumi Aramaki Shunsuke Furutani |
| author_sort | Izumi Kubo |
| collection | DOAJ |
| description | Salmonella <i>enterica</i> is a pathogenic bacterium that causes foodborne illness. One of the vehicle foods of <i>S. enterica</i> are chicken eggs. Efficient collection of the bacterium is necessary to detect it specifically. We developed a method to detect <i>S. enterica</i> by PCR on a microfluidic disc device using a fluorescent probe. Salmonella <i>enterica</i> cells were isolated in the microchambers on the device, followed by thermal lysis and PCR targeting with the invA gene, a gene specific to <i>S. enterica</i>, 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. <i>S. enterica</i> was detected at 5.0×10<sup>4</sup> cells mL<sup>−1</sup> or at a higher concentration of egg yolk within 6 h including the sampling time. |
| first_indexed | 2024-04-14T01:27:34Z |
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| id | doaj.art-bde512f41a2f41e4bc37f314a740fd0a |
| institution | Directory Open Access Journal |
| issn | 1424-8220 |
| language | English |
| last_indexed | 2024-04-14T01:27:34Z |
| publishDate | 2020-02-01 |
| publisher | MDPI AG |
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| series | Sensors |
| spelling | doaj.art-bde512f41a2f41e4bc37f314a740fd0a2022-12-22T02:20:21ZengMDPI AGSensors1424-82202020-02-01204106010.3390/s20041060s20041060Detection of Salmonella Enterica in Egg Yolk by PCR on a Microfluidic Disc Device Using Immunomagnetic BeadsIzumi Kubo0Mitsutoshi Kajiya1Narumi Aramaki2Shunsuke Furutani3Graduate School of Engineering, Soka University, 1-236 Tangi, Hachioji, Tokyo 192-8577, JapanGraduate School of Engineering, Soka University, 1-236 Tangi, Hachioji, Tokyo 192-8577, JapanGraduate School of Engineering, Soka University, 1-236 Tangi, Hachioji, Tokyo 192-8577, JapanBiomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-8-31 Midorigaoka, Ikeda, Osaka 563-8577, JapanSalmonella <i>enterica</i> is a pathogenic bacterium that causes foodborne illness. One of the vehicle foods of <i>S. enterica</i> are chicken eggs. Efficient collection of the bacterium is necessary to detect it specifically. We developed a method to detect <i>S. enterica</i> by PCR on a microfluidic disc device using a fluorescent probe. Salmonella <i>enterica</i> cells were isolated in the microchambers on the device, followed by thermal lysis and PCR targeting with the invA gene, a gene specific to <i>S. enterica</i>, 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. <i>S. enterica</i> was detected at 5.0×10<sup>4</sup> cells mL<sup>−1</sup> or at a higher concentration of egg yolk within 6 h including the sampling time.https://www.mdpi.com/1424-8220/20/4/1060immunomagnetic beadssalmonella <i>enterica</i>pcrinvaegg yolk |
| spellingShingle | Izumi Kubo Mitsutoshi Kajiya Narumi Aramaki Shunsuke Furutani Detection of Salmonella Enterica in Egg Yolk by PCR on a Microfluidic Disc Device Using Immunomagnetic Beads Sensors immunomagnetic beads salmonella <i>enterica</i> pcr inva egg yolk |
| title | Detection of Salmonella Enterica in Egg Yolk by PCR on a Microfluidic Disc Device Using Immunomagnetic Beads |
| title_full | Detection of Salmonella Enterica in Egg Yolk by PCR on a Microfluidic Disc Device Using Immunomagnetic Beads |
| title_fullStr | Detection of Salmonella Enterica in Egg Yolk by PCR on a Microfluidic Disc Device Using Immunomagnetic Beads |
| title_full_unstemmed | Detection of Salmonella Enterica in Egg Yolk by PCR on a Microfluidic Disc Device Using Immunomagnetic Beads |
| title_short | Detection of Salmonella Enterica in Egg Yolk by PCR on a Microfluidic Disc Device Using Immunomagnetic Beads |
| title_sort | detection of salmonella enterica in egg yolk by pcr on a microfluidic disc device using immunomagnetic beads |
| topic | immunomagnetic beads salmonella <i>enterica</i> pcr inva egg yolk |
| url | https://www.mdpi.com/1424-8220/20/4/1060 |
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