A universal fluorescence biosensor based on rolling circle amplification and locking probe for DNA detection.

Fang, Ying; Nie, Lanxin; Wang, Suqin; Liu, Shiwen; Li, Hongbo; Yu, Ruqin

Fang, Ying; Nie, Lanxin; Wang, Suqin; Liu, Shiwen; Li, Hongbo; Yu, Ruqin.

Afiliación

Fang Y; College of Chemistry and Chemical Engineering, Jiangxi Normal University, Nanchang, 330022, P.R. China.
Nie L; College of Chemistry and Chemical Engineering, Jiangxi Normal University, Nanchang, 330022, P.R. China.
Wang S; College of Chemistry and Chemical Engineering, Jiangxi Normal University, Nanchang, 330022, P.R. China.
Liu S; College of Chemistry and Chemical Engineering, Jiangxi Normal University, Nanchang, 330022, P.R. China. liushiwen985@126.com.
Li H; Jiangxi Provincial Center for Disease Control and Prevention, Nanchang, 330029, P. R. China. liushiwen985@126.com.
Yu R; College of Chemistry and Chemical Engineering, Jiangxi Normal University, Nanchang, 330022, P.R. China. lihongbo112@126.com.

Mikrochim Acta ; 191(7): 437, 2024 07 01.

Article en En | MEDLINE | ID: mdl-38951284

ABSTRACT

ABSTRACT

A stable DNA signal amplification sensor was developed on account of rolling circle amplification (RCA). This sensor includes target DNA-controlled rolling circle amplification technology and locking probe DNA replacement technology, which can be used to detect DNA fragments with genetic information, thus constructing a biosensor for universal detection of DNA. This study takes the homologous DNA of human immunodeficiency virus (HIV) and let-7a as examples to describe this biosensor. The padlock probe is first cyclized by T4 DNA ligase in response to the target's reaction with it. Then, rolling cycle amplification is initiated by Phi29 DNA polymerase, resulting in the formation of a lengthy chain with several triggers. These triggers can open the locked probe LP1 with the fluorescence signal turned off, so that it can continue to react with H2 to form a stable H1-H2 double strand. This regulates the distance between B-DNA modified by the quenching group and H1 modified by fluorescent group, and the fluorescence signal is recovered.

Asunto(s)

Técnicas Biosensibles; Sondas de ADN; Técnicas de Amplificación de Ácido Nucleico; Técnicas Biosensibles/métodos; Técnicas de Amplificación de Ácido Nucleico/métodos; Humanos; Sondas de ADN/química; Sondas de ADN/genética; Colorantes Fluorescentes/química; ADN Viral/análisis; ADN Viral/genética; ADN/química; ADN/genética; Espectrometría de Fluorescencia/métodos; Fluorescencia; ADN Polimerasa Dirigida por ADN/metabolismo; ADN Polimerasa Dirigida por ADN/química; Límite de Detección; VIH/genética

Palabras clave

Fluorescent detection; HIV; Homologous DNA of let-7a; Locking probe

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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Técnicas Biosensibles / Sondas de ADN / Técnicas de Amplificación de Ácido Nucleico Límite: Humans Idioma: En Revista: Mikrochim Acta Año: 2024 Tipo del documento: Article Pais de publicación: Austria

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