Electrochemical Method for Sulfamethazine Detection Based on CRISPR/Cas12a Coupled with Spherical Nucleic Acid with the Core of Gold Nanoparticle
Sulfamethazine was widely used to prevent or treat bacterial diseases in agricultural production such as livestock breeding and aquaculture. However, irrational use of sulfamethazine would lead to excessive residual amount in animal food, which would harm human health. Therefore, rapid and sensitive...
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The editorial department of Science and Technology of Food Industry
2025-04-01
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| Series: | Shipin gongye ke-ji |
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| Online Access: | http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2024060123 |
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| author | Yue HUANG Qian WANG Yan ZHANG Xianju QI Yaosong WANG |
| author_facet | Yue HUANG Qian WANG Yan ZHANG Xianju QI Yaosong WANG |
| author_sort | Yue HUANG |
| collection | DOAJ |
| description | Sulfamethazine was widely used to prevent or treat bacterial diseases in agricultural production such as livestock breeding and aquaculture. However, irrational use of sulfamethazine would lead to excessive residual amount in animal food, which would harm human health. Therefore, rapid and sensitive detection of sulfamethazine was of great significance for food safety and human health. In this work, a novel electrochemical method for the detection of sulfamethazine was constructed based on CRISPR/Cas12a coupled with spherical nucleic acid of gold nanoparticle core to realize rapid, highly sensitive and specific detection of sulfamethazine. The aptamer sequence of the target sulfamethazine was designed as the activator of CRISPR/Cas12a, and the specific binding of sulfamethazine to its aptamer sequence would inhibit the trans-cleavage activity of CRISPR/Cas12a, so that the substrate probe pre-modified on the surface of the gold electrode could be retained, and then the spherical nucleic acid could be introduced to trigger the hybridization chain reaction, producing significantly amplified electrochemical signal. To obtain the best analytical performance, several key conditions including the incubation time of sulfamethazine with aptamer, the concentration and cleavage time of CRISPR/Cas12a, the concentration and reaction time of hairpin probes were optimized, and the results were 30 min, 120 nmol/L, 30 min, 1 µmol/L, 100 min, respectively. Under the optimal conditions, the proposed electrochemical method exhibited prominent quantitative detection performance with the linear detection range of 0.006~600 ng/mL and the detection limit as low as 6 pg/mL. Meanwhile, the method showed good distinguishing ability between target antibiotic and other interfering antibiotics. In addition, the method was successfully applied to the detection of sulfamethazine in actual food samples with the recoveries ranging from 98.07%~106.90%, showing potential application value for food safety detection. |
| format | Article |
| id | doaj-art-32e1bd6fa6be431fa9537d73207f6c7e |
| institution | DOAJ |
| issn | 1002-0306 |
| language | zho |
| publishDate | 2025-04-01 |
| publisher | The editorial department of Science and Technology of Food Industry |
| record_format | Article |
| series | Shipin gongye ke-ji |
| spelling | doaj-art-32e1bd6fa6be431fa9537d73207f6c7e2025-08-20T03:09:11ZzhoThe editorial department of Science and Technology of Food IndustryShipin gongye ke-ji1002-03062025-04-0146830231010.13386/j.issn1002-0306.20240601232024060123-8Electrochemical Method for Sulfamethazine Detection Based on CRISPR/Cas12a Coupled with Spherical Nucleic Acid with the Core of Gold NanoparticleYue HUANG0Qian WANG1Yan ZHANG2Xianju QI3Yaosong WANG4College of Light Industry and Food Engineering, Nanjing Forestry University, Nanjing 210037, ChinaCollege of Light Industry and Food Engineering, Nanjing Forestry University, Nanjing 210037, ChinaCollege of Light Industry and Food Engineering, Nanjing Forestry University, Nanjing 210037, ChinaCollege of Light Industry and Food Engineering, Nanjing Forestry University, Nanjing 210037, ChinaCollege of Light Industry and Food Engineering, Nanjing Forestry University, Nanjing 210037, ChinaSulfamethazine was widely used to prevent or treat bacterial diseases in agricultural production such as livestock breeding and aquaculture. However, irrational use of sulfamethazine would lead to excessive residual amount in animal food, which would harm human health. Therefore, rapid and sensitive detection of sulfamethazine was of great significance for food safety and human health. In this work, a novel electrochemical method for the detection of sulfamethazine was constructed based on CRISPR/Cas12a coupled with spherical nucleic acid of gold nanoparticle core to realize rapid, highly sensitive and specific detection of sulfamethazine. The aptamer sequence of the target sulfamethazine was designed as the activator of CRISPR/Cas12a, and the specific binding of sulfamethazine to its aptamer sequence would inhibit the trans-cleavage activity of CRISPR/Cas12a, so that the substrate probe pre-modified on the surface of the gold electrode could be retained, and then the spherical nucleic acid could be introduced to trigger the hybridization chain reaction, producing significantly amplified electrochemical signal. To obtain the best analytical performance, several key conditions including the incubation time of sulfamethazine with aptamer, the concentration and cleavage time of CRISPR/Cas12a, the concentration and reaction time of hairpin probes were optimized, and the results were 30 min, 120 nmol/L, 30 min, 1 µmol/L, 100 min, respectively. Under the optimal conditions, the proposed electrochemical method exhibited prominent quantitative detection performance with the linear detection range of 0.006~600 ng/mL and the detection limit as low as 6 pg/mL. Meanwhile, the method showed good distinguishing ability between target antibiotic and other interfering antibiotics. In addition, the method was successfully applied to the detection of sulfamethazine in actual food samples with the recoveries ranging from 98.07%~106.90%, showing potential application value for food safety detection.http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2024060123crispr/cas12agold nanoparticlespherical nucleic acidsulfamethazineelectrochemical detection |
| spellingShingle | Yue HUANG Qian WANG Yan ZHANG Xianju QI Yaosong WANG Electrochemical Method for Sulfamethazine Detection Based on CRISPR/Cas12a Coupled with Spherical Nucleic Acid with the Core of Gold Nanoparticle Shipin gongye ke-ji crispr/cas12a gold nanoparticle spherical nucleic acid sulfamethazine electrochemical detection |
| title | Electrochemical Method for Sulfamethazine Detection Based on CRISPR/Cas12a Coupled with Spherical Nucleic Acid with the Core of Gold Nanoparticle |
| title_full | Electrochemical Method for Sulfamethazine Detection Based on CRISPR/Cas12a Coupled with Spherical Nucleic Acid with the Core of Gold Nanoparticle |
| title_fullStr | Electrochemical Method for Sulfamethazine Detection Based on CRISPR/Cas12a Coupled with Spherical Nucleic Acid with the Core of Gold Nanoparticle |
| title_full_unstemmed | Electrochemical Method for Sulfamethazine Detection Based on CRISPR/Cas12a Coupled with Spherical Nucleic Acid with the Core of Gold Nanoparticle |
| title_short | Electrochemical Method for Sulfamethazine Detection Based on CRISPR/Cas12a Coupled with Spherical Nucleic Acid with the Core of Gold Nanoparticle |
| title_sort | electrochemical method for sulfamethazine detection based on crispr cas12a coupled with spherical nucleic acid with the core of gold nanoparticle |
| topic | crispr/cas12a gold nanoparticle spherical nucleic acid sulfamethazine electrochemical detection |
| url | http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2024060123 |
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