Microfluidic Biosensor Integrated with Signal Transduction and Enhancement Mechanism for Ultrasensitive Noncompetitive Assay of Multiple Mycotoxins
To achieve high-throughput ultrasensitive detection of mycotoxins in food, a functional DNA-guided transition-state CRISPR/Cas12a microfluidic biosensor (named FTMB) was successfully constructed. The signal transduction CRISPR/Cas12a strategy in FTMB has utilized DNA sequences with a specific recogn...
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| Veröffentlicht in: | Analytical chemistry (Washington) 2023-05, Vol.95 (20), p.7993-8001 |
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| creator | Xiang, Xinran Song, Minghui Xu, Xiaowei Lu, Jiaran Chen, Yuanyuan Chen, Shuhan He, Yinglong Shang, Yuting |
| description | To achieve high-throughput ultrasensitive detection of mycotoxins in food, a functional DNA-guided transition-state CRISPR/Cas12a microfluidic biosensor (named FTMB) was successfully constructed. The signal transduction CRISPR/Cas12a strategy in FTMB has utilized DNA sequences with a specific recognition function and activators to form trigger switches. Meanwhile, the transition-state CRISPR/Cas12a system was constructed by adjusting the composition ratio of crRNA and activator to achieve a high response for low concentrations of target mycotoxins. On the other hand, the signal enhancement of FTMB has efficiently integrated the signal output of quantum dots (QDs) with the fluorescence enhancement effect of photonic crystals (PCs). The construction of universal QDs for the CRISPR/Cas12a system and PC films matching the photonic bandgap produced a significant signal enhancement by a factor of 45.6. Overall, FTMB exhibited a wide analytic range (10–5–101 ng·mL–1), low detection of limit (fg·mL–1), short detection period (∼40 min), high specificity, good precision (coefficients of variation |
| doi_str_mv | 10.1021/acs.analchem.3c00813 |
| format | Article |
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The signal transduction CRISPR/Cas12a strategy in FTMB has utilized DNA sequences with a specific recognition function and activators to form trigger switches. Meanwhile, the transition-state CRISPR/Cas12a system was constructed by adjusting the composition ratio of crRNA and activator to achieve a high response for low concentrations of target mycotoxins. On the other hand, the signal enhancement of FTMB has efficiently integrated the signal output of quantum dots (QDs) with the fluorescence enhancement effect of photonic crystals (PCs). The construction of universal QDs for the CRISPR/Cas12a system and PC films matching the photonic bandgap produced a significant signal enhancement by a factor of 45.6. Overall, FTMB exhibited a wide analytic range (10–5–101 ng·mL–1), low detection of limit (fg·mL–1), short detection period (∼40 min), high specificity, good precision (coefficients of variation <5%), and satisfactory practical sample analysis capacity (the consistency with HPLC at 88.76%–109.99%). It would provide a new and reliable solution for the rapid detection of multiple small molecules in the fields of clinical diagnosis and food safety.</description><identifier>ISSN: 0003-2700</identifier><identifier>EISSN: 1520-6882</identifier><identifier>DOI: 10.1021/acs.analchem.3c00813</identifier><identifier>PMID: 37156096</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject>Analytical chemistry ; Biological Assay ; Biosensing Techniques ; Biosensors ; Chemistry ; Chromatography, High Pressure Liquid ; Coefficient of variation ; CRISPR ; CRISPR-Cas Systems ; Crystals ; Deoxyribonucleic acid ; DNA ; Food safety ; Gene sequencing ; Liquid chromatography ; Low concentrations ; Mathematical analysis ; Microfluidics ; Mycotoxins ; Nucleotide sequence ; Photonic band gaps ; Photonic crystals ; Quantum dots ; Signal Transduction ; Switches</subject><ispartof>Analytical chemistry (Washington), 2023-05, Vol.95 (20), p.7993-8001</ispartof><rights>2023 American Chemical Society</rights><rights>Copyright American Chemical Society May 23, 2023</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a376t-9bd1c2b0237e9b464bf1e7d598dcedeb7f14c1ea42b40b76cb083ce0a42cc5343</citedby><cites>FETCH-LOGICAL-a376t-9bd1c2b0237e9b464bf1e7d598dcedeb7f14c1ea42b40b76cb083ce0a42cc5343</cites><orcidid>0000-0002-1139-9438</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/acs.analchem.3c00813$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/acs.analchem.3c00813$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>314,777,781,2752,27057,27905,27906,56719,56769</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/37156096$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Xiang, Xinran</creatorcontrib><creatorcontrib>Song, Minghui</creatorcontrib><creatorcontrib>Xu, Xiaowei</creatorcontrib><creatorcontrib>Lu, Jiaran</creatorcontrib><creatorcontrib>Chen, Yuanyuan</creatorcontrib><creatorcontrib>Chen, Shuhan</creatorcontrib><creatorcontrib>He, Yinglong</creatorcontrib><creatorcontrib>Shang, Yuting</creatorcontrib><title>Microfluidic Biosensor Integrated with Signal Transduction and Enhancement Mechanism for Ultrasensitive Noncompetitive Assay of Multiple Mycotoxins</title><title>Analytical chemistry (Washington)</title><addtitle>Anal. Chem</addtitle><description>To achieve high-throughput ultrasensitive detection of mycotoxins in food, a functional DNA-guided transition-state CRISPR/Cas12a microfluidic biosensor (named FTMB) was successfully constructed. The signal transduction CRISPR/Cas12a strategy in FTMB has utilized DNA sequences with a specific recognition function and activators to form trigger switches. Meanwhile, the transition-state CRISPR/Cas12a system was constructed by adjusting the composition ratio of crRNA and activator to achieve a high response for low concentrations of target mycotoxins. On the other hand, the signal enhancement of FTMB has efficiently integrated the signal output of quantum dots (QDs) with the fluorescence enhancement effect of photonic crystals (PCs). The construction of universal QDs for the CRISPR/Cas12a system and PC films matching the photonic bandgap produced a significant signal enhancement by a factor of 45.6. Overall, FTMB exhibited a wide analytic range (10–5–101 ng·mL–1), low detection of limit (fg·mL–1), short detection period (∼40 min), high specificity, good precision (coefficients of variation <5%), and satisfactory practical sample analysis capacity (the consistency with HPLC at 88.76%–109.99%). 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Chem</addtitle><date>2023-05-23</date><risdate>2023</risdate><volume>95</volume><issue>20</issue><spage>7993</spage><epage>8001</epage><pages>7993-8001</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><abstract>To achieve high-throughput ultrasensitive detection of mycotoxins in food, a functional DNA-guided transition-state CRISPR/Cas12a microfluidic biosensor (named FTMB) was successfully constructed. The signal transduction CRISPR/Cas12a strategy in FTMB has utilized DNA sequences with a specific recognition function and activators to form trigger switches. Meanwhile, the transition-state CRISPR/Cas12a system was constructed by adjusting the composition ratio of crRNA and activator to achieve a high response for low concentrations of target mycotoxins. On the other hand, the signal enhancement of FTMB has efficiently integrated the signal output of quantum dots (QDs) with the fluorescence enhancement effect of photonic crystals (PCs). The construction of universal QDs for the CRISPR/Cas12a system and PC films matching the photonic bandgap produced a significant signal enhancement by a factor of 45.6. Overall, FTMB exhibited a wide analytic range (10–5–101 ng·mL–1), low detection of limit (fg·mL–1), short detection period (∼40 min), high specificity, good precision (coefficients of variation <5%), and satisfactory practical sample analysis capacity (the consistency with HPLC at 88.76%–109.99%). It would provide a new and reliable solution for the rapid detection of multiple small molecules in the fields of clinical diagnosis and food safety.</abstract><cop>United States</cop><pub>American Chemical Society</pub><pmid>37156096</pmid><doi>10.1021/acs.analchem.3c00813</doi><tpages>9</tpages><orcidid>https://orcid.org/0000-0002-1139-9438</orcidid></addata></record> |
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| subjects | Analytical chemistry Biological Assay Biosensing Techniques Biosensors Chemistry Chromatography, High Pressure Liquid Coefficient of variation CRISPR CRISPR-Cas Systems Crystals Deoxyribonucleic acid DNA Food safety Gene sequencing Liquid chromatography Low concentrations Mathematical analysis Microfluidics Mycotoxins Nucleotide sequence Photonic band gaps Photonic crystals Quantum dots Signal Transduction Switches |
| title | Microfluidic Biosensor Integrated with Signal Transduction and Enhancement Mechanism for Ultrasensitive Noncompetitive Assay of Multiple Mycotoxins |
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