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...

Full description

Bibliographic Details
Main Authors: Chunxu Yu, Rujian Zhao, Yidan Tang, Siyan Zhou, Xin Zhao, Baiyang Lu, Fanzheng Meng, Bingling Li
Format: Article
Language:English
Published: KeAi Communications Co., Ltd. 2023-09-01
Series:Advanced Agrochem
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2773237123000436
_version_ 1797235493152227328
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
work_keys_str_mv AT chunxuyu functionalnucleicacidcircuitryasauniversaldetectorforpathogens
AT rujianzhao functionalnucleicacidcircuitryasauniversaldetectorforpathogens
AT yidantang functionalnucleicacidcircuitryasauniversaldetectorforpathogens
AT siyanzhou functionalnucleicacidcircuitryasauniversaldetectorforpathogens
AT xinzhao functionalnucleicacidcircuitryasauniversaldetectorforpathogens
AT baiyanglu functionalnucleicacidcircuitryasauniversaldetectorforpathogens
AT fanzhengmeng functionalnucleicacidcircuitryasauniversaldetectorforpathogens
AT binglingli functionalnucleicacidcircuitryasauniversaldetectorforpathogens