Single-Cell Detection of <i>Erwinia amylovora</i> Using Bio-Functionalized SIS Sensor
Herein, we developed a bio-functionalized solution-immersed silicon (SIS) sensor at the single-cell level to identify <i>Erwinia amylovora</i> (<i>E. amylovora</i>), a highly infectious bacterial pathogen responsible for fire blight, which is notorious for its rapid spread an...
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| Format: | Article |
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MDPI AG
2023-08-01
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| Series: | Sensors |
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| Online Access: | https://www.mdpi.com/1424-8220/23/17/7400 |
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| author | Ui Jin Lee Yunkwang Oh Oh Seok Kwon Jeong Mee Park Hyun Mo Cho Dong Hyung Kim Moonil Kim |
| author_facet | Ui Jin Lee Yunkwang Oh Oh Seok Kwon Jeong Mee Park Hyun Mo Cho Dong Hyung Kim Moonil Kim |
| author_sort | Ui Jin Lee |
| collection | DOAJ |
| description | Herein, we developed a bio-functionalized solution-immersed silicon (SIS) sensor at the single-cell level to identify <i>Erwinia amylovora</i> (<i>E. amylovora</i>), a highly infectious bacterial pathogen responsible for fire blight, which is notorious for its rapid spread and destructive impact on apple and pear orchards. This method allows for ultra-sensitive measurements without pre-amplification or labeling compared to conventional methods. To detect a single cell of <i>E. amylovora</i>, we used Lipopolysaccharide Transporter E (LptE), which is involved in the assembly of lipopolysaccharide (LPS) at the surface of the outer membrane of <i>E. amylovora</i>, as a capture agent. We confirmed that LptE interacts with <i>E. amylovora</i> via LPS through in-house ELISA analysis, then used it to construct the sensor chip by immobilizing the capture molecule on the sensor surface modified with 3′-Aminopropyl triethoxysilane (APTES) and glutaraldehyde (GA). The LptE-based SIS sensor exhibited the sensitive and specific detection of the target bacterial cell in real time. The dose–response curve shows a linearity (R<sup>2</sup> > 0.992) with wide dynamic ranges from 1 to 10<sup>7</sup> cells/mL for the target bacterial pathogen. The sensor showed the value change (d<i>Ψ</i>) of approximately 0.008° for growing overlayer thickness induced from a single-cell <i>E. amylovora</i>, while no change in the control bacterial cell (<i>Bacillus subtilis</i>) was observed, or negligible change, if any. Furthermore, the bacterial sensor demonstrated a potential for the continuous detection of <i>E. amylovora</i> through simple surface regeneration, enabling its reusability. Taken together, our system has the potential to be applied in fields where early symptoms are not observed and where single-cell or ultra-sensitive detection is required, such as plant bacterial pathogen detection, foodborne pathogen monitoring and analysis, and pathogenic microbial diagnosis. |
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| institution | Directory Open Access Journal |
| issn | 1424-8220 |
| language | English |
| last_indexed | 2024-03-10T23:13:17Z |
| publishDate | 2023-08-01 |
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| series | Sensors |
| spelling | doaj.art-562939cf11ee4028bab56372ad0a5a4f2023-11-19T08:49:22ZengMDPI AGSensors1424-82202023-08-012317740010.3390/s23177400Single-Cell Detection of <i>Erwinia amylovora</i> Using Bio-Functionalized SIS SensorUi Jin Lee0Yunkwang Oh1Oh Seok Kwon2Jeong Mee Park3Hyun Mo Cho4Dong Hyung Kim5Moonil Kim6Critical Diseases Diagnostics Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 125 Gwahang-ro, Yuseong-gu, Daejeon 34141, Republic of KoreaCritical Diseases Diagnostics Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 125 Gwahang-ro, Yuseong-gu, Daejeon 34141, Republic of KoreaSKKU Advanced Institute of Nanotechnology (SAINT), Sungkyunkwan University, Suwon 16419, Republic of KoreaPlant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 125 Gwahang-ro, Yuseong-gu, Daejeon 34141, Republic of KoreaDivision of Advanced Instrumentation Institute, Korea Research Institute of Standards and Science (KRISS), 267 Gajeong-ro, Yuseong-gu, Daejeon 34113, Republic of KoreaDivision of Advanced Instrumentation Institute, Korea Research Institute of Standards and Science (KRISS), 267 Gajeong-ro, Yuseong-gu, Daejeon 34113, Republic of KoreaCritical Diseases Diagnostics Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 125 Gwahang-ro, Yuseong-gu, Daejeon 34141, Republic of KoreaHerein, we developed a bio-functionalized solution-immersed silicon (SIS) sensor at the single-cell level to identify <i>Erwinia amylovora</i> (<i>E. amylovora</i>), a highly infectious bacterial pathogen responsible for fire blight, which is notorious for its rapid spread and destructive impact on apple and pear orchards. This method allows for ultra-sensitive measurements without pre-amplification or labeling compared to conventional methods. To detect a single cell of <i>E. amylovora</i>, we used Lipopolysaccharide Transporter E (LptE), which is involved in the assembly of lipopolysaccharide (LPS) at the surface of the outer membrane of <i>E. amylovora</i>, as a capture agent. We confirmed that LptE interacts with <i>E. amylovora</i> via LPS through in-house ELISA analysis, then used it to construct the sensor chip by immobilizing the capture molecule on the sensor surface modified with 3′-Aminopropyl triethoxysilane (APTES) and glutaraldehyde (GA). The LptE-based SIS sensor exhibited the sensitive and specific detection of the target bacterial cell in real time. The dose–response curve shows a linearity (R<sup>2</sup> > 0.992) with wide dynamic ranges from 1 to 10<sup>7</sup> cells/mL for the target bacterial pathogen. The sensor showed the value change (d<i>Ψ</i>) of approximately 0.008° for growing overlayer thickness induced from a single-cell <i>E. amylovora</i>, while no change in the control bacterial cell (<i>Bacillus subtilis</i>) was observed, or negligible change, if any. Furthermore, the bacterial sensor demonstrated a potential for the continuous detection of <i>E. amylovora</i> through simple surface regeneration, enabling its reusability. Taken together, our system has the potential to be applied in fields where early symptoms are not observed and where single-cell or ultra-sensitive detection is required, such as plant bacterial pathogen detection, foodborne pathogen monitoring and analysis, and pathogenic microbial diagnosis.https://www.mdpi.com/1424-8220/23/17/7400single-cell<i>Erwinia amylovora</i>fire blightLptESISbiosensor |
| spellingShingle | Ui Jin Lee Yunkwang Oh Oh Seok Kwon Jeong Mee Park Hyun Mo Cho Dong Hyung Kim Moonil Kim Single-Cell Detection of <i>Erwinia amylovora</i> Using Bio-Functionalized SIS Sensor Sensors single-cell <i>Erwinia amylovora</i> fire blight LptE SIS biosensor |
| title | Single-Cell Detection of <i>Erwinia amylovora</i> Using Bio-Functionalized SIS Sensor |
| title_full | Single-Cell Detection of <i>Erwinia amylovora</i> Using Bio-Functionalized SIS Sensor |
| title_fullStr | Single-Cell Detection of <i>Erwinia amylovora</i> Using Bio-Functionalized SIS Sensor |
| title_full_unstemmed | Single-Cell Detection of <i>Erwinia amylovora</i> Using Bio-Functionalized SIS Sensor |
| title_short | Single-Cell Detection of <i>Erwinia amylovora</i> Using Bio-Functionalized SIS Sensor |
| title_sort | single cell detection of i erwinia amylovora i using bio functionalized sis sensor |
| topic | single-cell <i>Erwinia amylovora</i> fire blight LptE SIS biosensor |
| url | https://www.mdpi.com/1424-8220/23/17/7400 |
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