One‐Step RAA and CRISPR‐Cas13a Method for Detecting Influenza B Virus DOI Creative Commons
Xinling Zhang, Shiyu Chen,

Juezhuo Li

et al.

Microbial Biotechnology, Journal Year: 2025, Volume and Issue: 18(4)

Published: April 1, 2025

ABSTRACT We developed a sensitive and specific method based on recombinase‐aided amplification (RAA) clustered regularly interspaced short palindromic repeats (CRISPR)‐CRISPR‐associated protein 13a (Cas13a). This method, named CRISPR‐based Rapid Efficient Test (CRISPRET), is designed for the early diagnosis of Influenza B (FluB) with aim shortening its transmission chain. identified conserved regions in Virus (IBV) NS gene forward reverse primers along crRNAs. then established optimised reaction system, Nucleic Acid Positive Reference Materials IBV were used to evaluate detection limit (DL) CRISPRET. Additionally, we collected 257 clinical samples, comprising 127 samples from patients infection 130 healthy individuals, subjected them dual using CRISPRET qPCR positive predictive value (PPV), negative (NPV), sensitivity specificity one primer, two primers, crRNAs establish optimise CRISPR ET. The demonstrated DL 500 copies·μL −1 when assisted by appropriate equipment. Despite requiring auxiliary equipment 30‐min reaction, ET enables nucleic acid within approximately first 5 min, achieving high (100%), (97.69%), PPV (97.69%) NPV concordance rate 98.83% qPCR. offers simple, field‐applicable, one‐step rapid IBV. It has strong potential field‐testing applications intelligent integration into existing diagnostic systems.

Language: Английский

Emerging microfluidic technologies for CRISPR-based diagnostics: an overview DOI
Fatemeh Nafian, Kimia Sadat Esfahani, Mina Hobabi Aghmiuni

et al.

Analytical Methods, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 1, 2025

This review explores emerging CRISPR-microfluidic platforms enhancing precision, speed, and portability for point-of-care diagnostics. Innovations like SHINE, CARMEN, ITP, DNAiTECH, Dμchip, FAST, MAPnavi highlight their potential.

Language: Английский

Citations

0
Simultaneous detection of Enterovirus 71 and Coxsackievirus A16 based on RT-RRA-CRISPR Cas12a/13a system DOI
Zhiyun Wu,

Mengyuan Tang,

Xue Li

et al.

Microchemical Journal, Journal Year: 2025, Volume and Issue: unknown, P. 113335 - 113335

Published: March 1, 2025

Language: Английский

Citations

0
One‐Step RAA and CRISPR‐Cas13a Method for Detecting Influenza B Virus DOI Creative Commons
Xinling Zhang, Shiyu Chen,

Juezhuo Li

et al.

Microbial Biotechnology, Journal Year: 2025, Volume and Issue: 18(4)

Published: April 1, 2025

ABSTRACT We developed a sensitive and specific method based on recombinase‐aided amplification (RAA) clustered regularly interspaced short palindromic repeats (CRISPR)‐CRISPR‐associated protein 13a (Cas13a). This method, named CRISPR‐based Rapid Efficient Test (CRISPRET), is designed for the early diagnosis of Influenza B (FluB) with aim shortening its transmission chain. identified conserved regions in Virus (IBV) NS gene forward reverse primers along crRNAs. then established optimised reaction system, Nucleic Acid Positive Reference Materials IBV were used to evaluate detection limit (DL) CRISPRET. Additionally, we collected 257 clinical samples, comprising 127 samples from patients infection 130 healthy individuals, subjected them dual using CRISPRET qPCR positive predictive value (PPV), negative (NPV), sensitivity specificity one primer, two primers, crRNAs establish optimise CRISPR ET. The demonstrated DL 500 copies·μL −1 when assisted by appropriate equipment. Despite requiring auxiliary equipment 30‐min reaction, ET enables nucleic acid within approximately first 5 min, achieving high (100%), (97.69%), PPV (97.69%) NPV concordance rate 98.83% qPCR. offers simple, field‐applicable, one‐step rapid IBV. It has strong potential field‐testing applications intelligent integration into existing diagnostic systems.

Language: Английский

Citations

0