Naked-eye detection with loop-mediated isothermal amplification for P. carotovorum subsp. carotovorum in agricultural products
Pectobacterium carotovorum causing soft-rot disease requires on-site detection before the distribution of agricultural products. Loop-mediated isothermal amplification (LAMP), which is resistant to food inhibitors, is known for its high detection sensitivity for pathogens and when coupled with later...
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| Published in | Food science and biotechnology Vol. 33; no. 1; pp. 203 - 209 |
|---|---|
| Main Authors | , , , |
| Format | Journal Article |
| Language | English |
| Published |
Singapore
Springer Nature Singapore
01.01.2024
Springer Nature B.V 한국식품과학회 |
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| Abstract | Pectobacterium carotovorum
causing soft-rot disease requires on-site detection before the distribution of agricultural products. Loop-mediated isothermal amplification (LAMP), which is resistant to food inhibitors, is known for its high detection sensitivity for pathogens and when coupled with lateral flow immunoassay (LFA) enables visualizations. For detection of soft-rot disease, we developed a LAMP-LFA system targeting 16S ribosomal RNA, a partial sequence gene of
P. carotovorum
subsp.
carotovorum
. The LAMP-LFA was performed at 60 °C for 50 min followed by hybridization of digoxygenin-labeled LAMP amplicon and biotinylated probe. Detection sensitivity was 3.22 × 10
1
CFU/mL in pure culture, which specifically detected the target. In Chinese cabbage and potato, the target was detected up to low levels of 1.57 × 10
2
CFU/g and 1.29 × 10
2
CFU/g, respectively. This study showed potential applicability as a sensitive point-of-care system for soft-rot disease bacteria detection in agricultural products. |
|---|---|
| AbstractList | causing soft-rot disease requires on-site detection before the distribution of agricultural products. Loop-mediated isothermal amplification (LAMP), which is resistant to food inhibitors, is known for its high detection sensitivity for pathogens and when coupled with lateral flow immunoassay (LFA) enables visualizations. For detection of soft-rot disease, we developed a LAMP-LFA system targeting 16S ribosomal RNA, a partial sequence gene of
subsp.
. The LAMP-LFA was performed at 60 °C for 50 min followed by hybridization of digoxygenin-labeled LAMP amplicon and biotinylated probe. Detection sensitivity was 3.22 × 10
CFU/mL in pure culture, which specifically detected the target. In Chinese cabbage and potato, the target was detected up to low levels of 1.57 × 10
CFU/g and 1.29 × 10
CFU/g, respectively. This study showed potential applicability as a sensitive point-of-care system for soft-rot disease bacteria detection in agricultural products.
The online version contains supplementary material available at 10.1007/s10068-023-01315-z. Pectobacterium carotovorum causing soft-rot disease requires on-site detection before the distribution of agricultural products. Loop-mediated isothermal amplification (LAMP), which is resistant to food inhibitors, is known for its high detection sensitivity for pathogens and when coupled with lateral flow immunoassay (LFA) enables visualizations. For detection of soft-rot disease, we developed a LAMP-LFA system targeting 16S ribosomal RNA, a partial sequence gene of P. carotovorum subsp. carotovorum . The LAMP-LFA was performed at 60 °C for 50 min followed by hybridization of digoxygenin-labeled LAMP amplicon and biotinylated probe. Detection sensitivity was 3.22 × 10 1 CFU/mL in pure culture, which specifically detected the target. In Chinese cabbage and potato, the target was detected up to low levels of 1.57 × 10 2 CFU/g and 1.29 × 10 2 CFU/g, respectively. This study showed potential applicability as a sensitive point-of-care system for soft-rot disease bacteria detection in agricultural products. Pectobacterium carotovorum causing soft-rot disease requires on-site detection before the distribution of agricultural products. Loop-mediated isothermal amplification (LAMP), which is resistant to food inhibitors, is known for its high detection sensitivity for pathogens and when coupled with lateral flow immunoassay (LFA) enables visualizations. For detection of soft-rot disease, we developed a LAMP-LFA system targeting 16S ribosomal RNA, a partial sequence gene of P. carotovorum subsp. carotovorum. The LAMP-LFA was performed at 60 °C for 50 min followed by hybridization of digoxygenin-labeled LAMP amplicon and biotinylated probe. Detection sensitivity was 3.22 × 101 CFU/mL in pure culture, which specifically detected the target. In Chinese cabbage and potato, the target was detected up to low levels of 1.57 × 102 CFU/g and 1.29 × 102 CFU/g, respectively. This study showed potential applicability as a sensitive point-of-care system for soft-rot disease bacteria detection in agricultural products.Supplementary InformationThe online version contains supplementary material available at 10.1007/s10068-023-01315-z. Pectobacterium carotovorum causing soft-rot disease requires on-site detection before the distribution of agricultural products. Loop-mediated isothermal amplification (LAMP), which is resistant to food inhibitors, is known for its high detection sensitivity for pathogens and when coupled with lateral flow immunoassay (LFA) enables visualizations. For detection of soft-rot disease, we developed a LAMP-LFA system targeting 16S ribosomal RNA, a partial sequence gene of P. carotovorum subsp. carotovorum. The LAMP-LFA was performed at 60 °C for 50 min followed by hybridization of digoxygenin-labeled LAMP amplicon and biotinylated probe. Detection sensitivity was 3.22 × 101 CFU/mL in pure culture, which specifically detected the target. In Chinese cabbage and potato, the target was detected up to low levels of 1.57 × 102 CFU/g and 1.29 × 102 CFU/g, respectively. This study showed potential applicability as a sensitive point-of-care system for soft-rot disease bacteria detection in agricultural products. KCI Citation Count: 0 Pectobacterium carotovorum causing soft-rot disease requires on-site detection before the distribution of agricultural products. Loop-mediated isothermal amplification (LAMP), which is resistant to food inhibitors, is known for its high detection sensitivity for pathogens and when coupled with lateral flow immunoassay (LFA) enables visualizations. For detection of soft-rot disease, we developed a LAMP-LFA system targeting 16S ribosomal RNA, a partial sequence gene of P. carotovorum subsp. carotovorum. The LAMP-LFA was performed at 60 °C for 50 min followed by hybridization of digoxygenin-labeled LAMP amplicon and biotinylated probe. Detection sensitivity was 3.22 × 101 CFU/mL in pure culture, which specifically detected the target. In Chinese cabbage and potato, the target was detected up to low levels of 1.57 × 102 CFU/g and 1.29 × 102 CFU/g, respectively. This study showed potential applicability as a sensitive point-of-care system for soft-rot disease bacteria detection in agricultural products. |
| Author | Oh, Se-Wook Moon, Ye-Ji Lee, So-Young Kim, Unji |
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| Cites_doi | 10.1016/j.micpath.2017.03.016 10.1038/s41598-021-81312-6 10.1007/s00604-017-2562-z 10.1111/lam.12590 10.1021/acs.jafc.9b06979 10.1038/s41598-021-01196-4 10.1093/nar/28.12.e63 10.1146/annurev-phyto-080417-045906 10.1016/j.foodchem.2020.126945 10.1016/j.lwt.2021.112242 10.1080/00219266.2008.9656148 10.1016/j.mcp.2017.05.004 10.1007/978-1-61779-609-8_13 10.1016/j.bcab.2016.11.004 10.3390/foods11030322 10.1007/s13197-019-03740-7 10.1099/ijsem.0.001485 10.1016/j.hpj.2020.07.004 10.1016/j.biosystemseng.2018.01.001 10.1021/acsnano.0c10035 10.9734/arrb/2020/v35i830257 10.1021/acs.jafc.8b01765 10.1002/anie.201109115 10.3389/fpls.2022.855262 10.1016/j.aca.2022.339781 10.3390/mi8090266 10.1016/j.foodcont.2019.04.014 10.1111/mpp.12378 10.1021/acs.jafc.7b03655 10.1016/j.micres.2022.127072 |
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| Keywords | Loop-mediated isothermal amplification Agricultural product Soft-rot disease Visualization detection Lateral flow immunoassay Pectobacterium carotovorum |
| Language | English |
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| Snippet | Pectobacterium carotovorum
causing soft-rot disease requires on-site detection before the distribution of agricultural products. Loop-mediated isothermal... causing soft-rot disease requires on-site detection before the distribution of agricultural products. Loop-mediated isothermal amplification (LAMP), which is... Pectobacterium carotovorum causing soft-rot disease requires on-site detection before the distribution of agricultural products. Loop-mediated isothermal... |
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| SubjectTerms | Agricultural products Chemistry Chemistry and Materials Science Chinese cabbage Food Science Gene amplification Hybridization Immunoassay Nucleotide sequence Nutrition Potatoes Pure culture Research Article Rot rRNA 16S Sensitivity Target detection 식품과학 |
| Title | Naked-eye detection with loop-mediated isothermal amplification for P. carotovorum subsp. carotovorum in agricultural products |
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