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Chemistry ; 29(27): e202300075, 2023 May 11.
Artículo en Inglés | MEDLINE | ID: mdl-36790320

RESUMEN

A new method for the detection of genomic RNA combines RNA cleavage by the 10-23 DNAzyme and use of the cleavage fragments as primers to initiate rolling circle amplification (RCA). 230 different 10-23 DNAzyme variants were screened to identify those that target accessible RNA sites within the highly structured RNA transcripts of SARS-CoV-2. A total of 28 DNAzymes were identified with >20 % cleavage, 5 with >40 % cleavage and one with >60 % in 10 min. The cleavage fragments from these reactions were then screened for coupling to an RCA reaction, leading to the identification of several cleavage fragments that could efficiently initiate RCA. Using a newly developed quasi-exponential RCA method with a detection limit of 500 aM of RNA, 14 RT-PCR positive and 15 RT-PCR negative patient saliva samples were evaluated for SARS-CoV-2 genomic RNA, achieving a clinical sensitivity of 86 % and specificity of 100 % for detection of the virus in <2.5 h.


Asunto(s)
Técnicas Biosensibles , COVID-19 , ADN Catalítico , Humanos , ADN Catalítico/metabolismo , ARN , SARS-CoV-2/genética , SARS-CoV-2/metabolismo , División del ARN , COVID-19/diagnóstico , Técnicas de Amplificación de Ácido Nucleico/métodos , Genómica , Técnicas Biosensibles/métodos
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