Naked-eye detection of site-specific ssRNA and ssDNA using PAMmer-assisted CRISPR/Cas9 coupling with exponential amplification reaction

One-Pot Visual Detection of African Swine Fever Virus Using CRISPR-Cas12a

“…The Cas12a enzyme itself has weak collateral cleavage activity; thus, it can only achieve low detection sensitivity without pre-amplification ( 27 , 58 ). Therefore, in combination with isothermal amplification, such as RPA, LAMP, strand displacement amplification (SDA) ( 59 – 61 ), rolling circle amplification (RCA) ( 62 , 63 ), exponential amplification reaction (EXPAR) ( 64 , 65 ), and recombinase-aided amplification (RAA) ( 48 , 66 – 68 ), we found that Cas12a was able to detect pathogens with high sensitivity and precision. In this study, we found that the CRISPR-RPA system is not highly sensitive, but it is faster than CRISPR-LAMP, which means that CRISPR-RPA is possible for rapid qualitative detection on the grassroots level.…”

CRISPR/Cas Systems‐Inspired Nano/Biosensors for Detecting Infectious Viruses and Pathogenic Bacteria

“…[84][85][86][87][88] Enzyme/nanozyme-catalyzed colorimetric sensing is established mainly based on the enzymes/nanozymes catalyzing the chromogenic substrates like 3,3',5,5'-tetramethylbenzidine (TMB) and OPD to generate the visible colorimetric output signals. [58,64,[89][90][91][92] Intriguingly, CRISPR/Cas systems have been introduced into the enzyme/ nanozyme-catalyzed colorimetric sensing for the construction of a novel detection platform. [93][94][95][96][97][98][99] As reported by Zhu's group, CRISPR/dCas9 system, split-horseradish peroxidase (HRP) techniques, isothermal amplification, and rolling circle amplification (RCA) were used to establish a new and lowcost RCA-CRISPR-split-HRP (RCH) strategy for the highly efficient detection of miRNAs with a single-base specificity.…”

Powerful CRISPR-Based Biosensing Techniques and Their Integration With Microfluidic Platforms

“…With these advantages, CRISPR/Cas12a- or CRISPR/Cas13a-triggered RCA is developed as a highly sensitive and specific biosensor, for example, for the detection of microRNAs ( Li D. et al, 2020 ; Tian et al, 2020 ; Qing et al, 2021 ; Zhou et al, 2021 ). Alternative isothermal amplification approaches, such as exponential amplification reaction (EXPAR) ( Song J. et al, 2021 ; Wang et al, 2021b ), hybridization chain reaction (HCR) ( Xing et al, 2020 ; Kachwala et al, 2021 ; Liu et al, 2022 ), and strand displacement amplification (SDA) ( Chen et al, 2021b ; Deng et al, 2021 ; Gong et al, 2021 ) were combined with CRISPR systems for sensing pathogenic bacteria and viruses, gene mutation, and even proteins. Different characteristics of this isothermal amplification method were compared as listed in Table 2 .…”