Article ; Online: Carrying out pseudo dual nucleic acid detection from sample to visual result in a polypropylene bag with CRISPR/Cas12a.
2021 Volume 178, Page(s) 113001
Abstract: ... environment monitoring. However, conventional nucleic acid analysis has to be carried out in laboratories ... by themselves, the application of nucleic acid detection would be greatly accelerated. We herein reported a polypropylene (PP ... bag-based method for convenient detection of nucleic acids in the oil-sealed space. The PP bag has ...
| Abstract | Amplification-based nucleic acid detection is widely employed in food safety, medical diagnosis and environment monitoring. However, conventional nucleic acid analysis has to be carried out in laboratories because of requiring expensive instruments and trained personnel. If people could do nucleic acid detection at home by themselves, the application of nucleic acid detection would be greatly accelerated. We herein reported a polypropylene (PP) bag-based method for convenient detection of nucleic acids in the oil-sealed space. The PP bag has three chambers which are responsible for lysis, washing and amplification/detection, respectively. After adding sample, nucleic acids are adsorbed on magnetic particles (MPs) and moved into these three chambers successively through immiscible oil channel by an external magnet. Combined with isothermal amplification, the PP bag can be incubated in a water bath or milk warmer and acted as a reaction tube. With highly specific CRISPR technology, Salmonella typhimurium (St) and SARS-CoV-2 can be visually detected in these PP bags within 1 h, indicating its potential household application. To further improve the reliability of nucleic acid testing at home, a logic decision method is introduced by detecting both target and endogenous reference gene. Positive/negative/invalid detection result can be obtained by chronologically adding the CRISPR reagents of target and endogenous reference gene. We anticipate that this PP bag can provide a novel toolkit for nucleic acid detection in people's daily life. |
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| MeSH term(s) | Biosensing Techniques/instrumentation ; Biosensing Techniques/methods ; COVID-19/diagnosis ; COVID-19/virology ; COVID-19 Nucleic Acid Testing/instrumentation ; COVID-19 Nucleic Acid Testing/methods ; CRISPR-Cas Systems ; DNA, Bacterial/genetics ; DNA, Bacterial/isolation & purification ; Food Microbiology ; Humans ; Magnetics ; Molecular Diagnostic Techniques/instrumentation ; Molecular Diagnostic Techniques/methods ; Nucleic Acid Amplification Techniques/instrumentation ; Nucleic Acid Amplification Techniques/methods ; Polypropylenes ; RNA, Viral/genetics ; RNA, Viral/isolation & purification ; SARS-CoV-2/genetics ; SARS-CoV-2/isolation & purification ; Salmonella typhimurium/genetics ; Salmonella typhimurium/isolation & purification ; Self-Testing | |||||
| Chemical Substances | DNA, Bacterial ; Polypropylenes ; RNA, Viral | |||||
| Language | English | |||||
| Publishing date | 2021-01-19 | |||||
| Publishing country | England | |||||
| Document type | Journal Article | |||||
| ZDB-ID | 1011023-9 | |||||
| ISSN | 1873-4235 ; 0956-5663 | |||||
| ISSN (online) | 1873-4235 | |||||
| ISSN | 0956-5663 | |||||
| DOI | 10.1016/j.bios.2021.113001 | |||||
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| Database | MEDical Literature Analysis and Retrieval System OnLINE |
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