Functional nucleic acid circuitry as a universal detector for pathogens
While the pathogen nucleic acid diagnostic technology has made tremendous progresses, several challenges, including long development cycles and limited specificity still exist, especially in the context of isothermal amplification techniques. To enhance the detection accuracy, here a functional stra...
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Format: | Article |
Language: | English |
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KeAi Communications Co., Ltd.
2023-09-01
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Series: | Advanced Agrochem |
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Online Access: | http://www.sciencedirect.com/science/article/pii/S2773237123000436 |
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author | Chunxu Yu Rujian Zhao Yidan Tang Siyan Zhou Xin Zhao Baiyang Lu Fanzheng Meng Bingling Li |
author_facet | Chunxu Yu Rujian Zhao Yidan Tang Siyan Zhou Xin Zhao Baiyang Lu Fanzheng Meng Bingling Li |
author_sort | Chunxu Yu |
collection | DOAJ |
description | While the pathogen nucleic acid diagnostic technology has made tremendous progresses, several challenges, including long development cycles and limited specificity still exist, especially in the context of isothermal amplification techniques. To enhance the detection accuracy, here a functional strand displacement catalytic hairpin assembly circuit, which can perform at high-temperature (HT-CHA), was developed as the downstream of the loop mediated isothermal nucleic acid amplification (LAMP). The addition of HT-CHA not only ensures the specificity but also amplifies the detection signal. Taking African swine fever (ASF) gene as the target, the input of HT-CHA was designed according to the ASF gene LAMP amplicons. This LAMP-HTCHA can detect as low as 2 copies/μL targeting genes with high signal-to-noise ratio. Through importing a three-way junction (3WJ) transducer, the HT-CHA well-developed for ASF gene can be directly adapted to detect the LAMP amplicons of other pathogen genes, such as Mycoplasma pneumoniae (MP) gene. The time-consuming and high-risk process to redesign HT-CHA components can be further avoided, making the method even general and useful for a plenty of other targets. Finally, the accurate detection of MP gene in alveolar lavage fluid samples confirmed the high potential of the LAMP and HT-CHA combination in clinical applications, providing a promising strategy to develop point-of-care diagnostics at constant temperatures. |
first_indexed | 2024-04-24T16:48:50Z |
format | Article |
id | doaj.art-ce925e67c26641c98309304021a12431 |
institution | Directory Open Access Journal |
issn | 2773-2371 |
language | English |
last_indexed | 2024-04-24T16:48:50Z |
publishDate | 2023-09-01 |
publisher | KeAi Communications Co., Ltd. |
record_format | Article |
series | Advanced Agrochem |
spelling | doaj.art-ce925e67c26641c98309304021a124312024-03-29T05:51:40ZengKeAi Communications Co., Ltd.Advanced Agrochem2773-23712023-09-0123291296Functional nucleic acid circuitry as a universal detector for pathogensChunxu Yu0Rujian Zhao1Yidan Tang2Siyan Zhou3Xin Zhao4Baiyang Lu5Fanzheng Meng6Bingling Li7State Key Lab of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China; School of Applied Chemistry and Engineering, University of Science and Technology of China, Hefei 230026, ChinaState Key Lab of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China; School of Applied Chemistry and Engineering, University of Science and Technology of China, Hefei 230026, ChinaState Key Lab of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, ChinaCenter for Pathogen Biology and Infectious Diseases, First Hospital of Jilin University, Changchun, 130021, ChinaCenter for Pathogen Biology and Infectious Diseases, First Hospital of Jilin University, Changchun, 130021, ChinaState Key Lab of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China; Corresponding author.Center for Pathogen Biology and Infectious Diseases, First Hospital of Jilin University, Changchun, 130021, China; Corresponding author.State Key Lab of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China; School of Applied Chemistry and Engineering, University of Science and Technology of China, Hefei 230026, China; Corresponding author.State Key Lab of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China.While the pathogen nucleic acid diagnostic technology has made tremendous progresses, several challenges, including long development cycles and limited specificity still exist, especially in the context of isothermal amplification techniques. To enhance the detection accuracy, here a functional strand displacement catalytic hairpin assembly circuit, which can perform at high-temperature (HT-CHA), was developed as the downstream of the loop mediated isothermal nucleic acid amplification (LAMP). The addition of HT-CHA not only ensures the specificity but also amplifies the detection signal. Taking African swine fever (ASF) gene as the target, the input of HT-CHA was designed according to the ASF gene LAMP amplicons. This LAMP-HTCHA can detect as low as 2 copies/μL targeting genes with high signal-to-noise ratio. Through importing a three-way junction (3WJ) transducer, the HT-CHA well-developed for ASF gene can be directly adapted to detect the LAMP amplicons of other pathogen genes, such as Mycoplasma pneumoniae (MP) gene. The time-consuming and high-risk process to redesign HT-CHA components can be further avoided, making the method even general and useful for a plenty of other targets. Finally, the accurate detection of MP gene in alveolar lavage fluid samples confirmed the high potential of the LAMP and HT-CHA combination in clinical applications, providing a promising strategy to develop point-of-care diagnostics at constant temperatures.http://www.sciencedirect.com/science/article/pii/S2773237123000436catalytic hairpin assemblyLoop-mediated isothermal amplificationThree-way junctionUniversalityPathogen detection |
spellingShingle | Chunxu Yu Rujian Zhao Yidan Tang Siyan Zhou Xin Zhao Baiyang Lu Fanzheng Meng Bingling Li Functional nucleic acid circuitry as a universal detector for pathogens Advanced Agrochem catalytic hairpin assembly Loop-mediated isothermal amplification Three-way junction Universality Pathogen detection |
title | Functional nucleic acid circuitry as a universal detector for pathogens |
title_full | Functional nucleic acid circuitry as a universal detector for pathogens |
title_fullStr | Functional nucleic acid circuitry as a universal detector for pathogens |
title_full_unstemmed | Functional nucleic acid circuitry as a universal detector for pathogens |
title_short | Functional nucleic acid circuitry as a universal detector for pathogens |
title_sort | functional nucleic acid circuitry as a universal detector for pathogens |
topic | catalytic hairpin assembly Loop-mediated isothermal amplification Three-way junction Universality Pathogen detection |
url | http://www.sciencedirect.com/science/article/pii/S2773237123000436 |
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