Cas12a-based electrochemiluminescence biosensor for target amplification-free DNA detection

Liu, Pengfei; Zhao, Kai-Ren; Liu, Zhijun; Wang, Li; Ye, Shu-Ying; Liang, Guoxi

doi:10.1016/j.bios.2020.112954

Cited by 106 publications

(57 citation statements)

References 49 publications

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“…Then, with the employment of Cas12 and Cas13, SHERLOCKv2 was created to simultaneously detect multiple target nucleic acids in one diagnostic system ( Gootenberg et al, 2018 ). Recently, with the use of either tandem crRNAs or remarkably reduced reaction volumes for target nucleic acid detection, diagnostic sensitivities can be greatly improved, and several amplification-free systems have been successfully developed ( Fozouni et al, 2021 ; Liu et al, 2021 ; Shinoda et al, 2021 ; Tian et al, 2021 ; Yue et al, 2021 ).…”

Section: Introductionmentioning

confidence: 99%

Definition of CRISPR Cas12a Trans-Cleavage Units to Facilitate CRISPR Diagnostics

Wang

Zhang

et al. 2021

Front. Microbiol.

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The CRISPR diagnostic (CRISPR-Dx) technology that employs the trans-cleavage activities has shown great potential in diagnostic sensitivity, specificity, convenience, and portability, and has been recognized as the next-generation diagnostic methods. However, due to the lack of standardized definition of Cas trans-cleavage enzymatic units, it is difficult to standardize the present CRISPR-Dx systems, which have undoubtedly impeded the development of the CRISPR-Dx industry. To solve the problem, we here first systematically optimized the reaction systems for Cas12a, and then defined its trans-cleavage units (transU), which we believe will be of great importance and interest to researchers in both molecular diagnostic industry and basic research. Moreover, a simple protocol was provided to facilitate a step-by-step measurement of the Cas12a transU, which can also act as a reference for the definition of the transU for other Cas proteins.

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Section: Introductionmentioning

confidence: 99%

Definition of CRISPR Cas12a Trans-Cleavage Units to Facilitate CRISPR Diagnostics

Wang

Zhang

et al. 2021

Front. Microbiol.

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“…Recently, it was shown that the amplification step may be avoided through signal amplification. One such option is to use gold nanoparticles, or L-methionine gold nanoclusters combined with electrochemiluminescence to enhance and stabilize the signal [ 52 , 53 ]. Another approach was recently reported where the use of several crRNAs, targeting several loci within the target DNA, significantly increased the fluorescent signal achieved following Cas13 activation [ 54 ].…”

Section: Discussionmentioning

confidence: 99%

Differential Detection of the Tobamoviruses Tomato Mosaic Virus (ToMV) and Tomato Brown Rugose Fruit Virus (ToBRFV) Using CRISPR-Cas12a

Alon

Hak

Bornstein

et al. 2021

Plants

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CRISPR/Cas12a-based detection is a novel approach for the efficient, sequence-specific identification of viruses. Here we adopt the use of CRISPR/Cas12a to identify the tomato brown rugose fruit virus (ToBRFV), a new and emerging tobamovirus which is causing substantial damage to the global tomato industry. Specific CRISPR RNAs (crRNAs) were designed to detect either ToBRFV or the closely related tomato mosaic virus (ToMV). This technology enabled the differential detection of ToBRFV and ToMV. Sensitivity assays revealed that viruses can be detected from 15–30 ng of RT-PCR product, and that specific detection could be achieved from a mix of ToMV and ToBRFV. In addition, we show that this method can enable the identification of ToBRFV in samples collected from commercial greenhouses. These results demonstrate a new method for species-specific detection of tobamoviruses. A future combination of this approach with isothermal amplification could provide a platform for efficient and user-friendly ways to distinguish between closely related strains and resistance-breaking pathogens.

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“…Electrochemiluminescence (ECL) biosensors for the detection of viruses, namely HIV and human papilloma virus (HPV) have been reported [64] , [65] , [66] . Zhao et al integrated spherical nucleic acid (SNA) with CRISPR/Cas12a for the detection of HIV and HPV [64] .…”

Section: Biosensors For the Detection Of Pathogensmentioning

confidence: 99%

“…Thus, it is recommended that further studied be made in efforts to simplify such detection schemes. Liu et al reported the development of a similar ECL biosensor using CRISPR/Cas for the detection of HPV-16 DNA [65] . Their biosensor was found to exhibit an LOD of 0.00048 nM.…”

Section: Biosensors For the Detection Of Pathogensmentioning

confidence: 99%

Recent advances in the biosensors application for the detection of bacteria and viruses in wastewater

Kadadou¹,

Tizani²,

Wadi³

et al. 2022

Journal of Environmental Chemical Engineering

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Cas12a-based electrochemiluminescence biosensor for target amplification-free DNA detection

Cited by 106 publications

References 49 publications

Definition of CRISPR Cas12a Trans-Cleavage Units to Facilitate CRISPR Diagnostics

Definition of CRISPR Cas12a Trans-Cleavage Units to Facilitate CRISPR Diagnostics

Differential Detection of the Tobamoviruses Tomato Mosaic Virus (ToMV) and Tomato Brown Rugose Fruit Virus (ToBRFV) Using CRISPR-Cas12a

Recent advances in the biosensors application for the detection of bacteria and viruses in wastewater

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