CRISPR-Cas12a-based detection of monkeypox virus

Sui, Yutong; Xu, Qi; Liu, Mingsheng; Zuo, Kuiyang; Liu, Xiaomei; Liu, Jinyu

doi:10.1016/j.jinf.2022.08.043

Cited by 16 publications

(20 citation statements)

References 4 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Previous report on CRISPR-based monkeypox detection lack the ability to differentiate it from other poxviruses [19]. Therefore, an attempt was made to distinguish the monkeypox virus from other poxviruses by leveraging presence of a monkeypox-specific SNP in polA gene.…”

Section: Conclusion and Discussionmentioning

confidence: 99%

“…This is well suited for detection in remote areas as expensive instrumentation such as RT-PCR machine is not required. Further, the assay is also compatible with the CRISPR-CUBE [11], a highly portable low-cost battery-operated incubator-cum-detector, in which could be an advantage over previous CRISPR-based monkeypox detection reports [19], [31]. Use of CRISPR-CUBE eliminates the recurring cost of lateral flow strips.…”

Section: Conclusion and Discussionmentioning

confidence: 99%

See 1 more Smart Citation

CRISPR/Cas12a assisted specific detection of monkeypox virus

Singh

Misra

Bindal

et al. 2022

Preprint

View full text Add to dashboard Cite

ObjectiveTo identify monkeypox-specific sequence and distinguish it from related orthopoxviruses followed by development of a CRISPR-Cas12a-based, specific and sensitive detection of monkeypox virus.MethodsA common detection mixture (CDM) was constituted comprising of CRISPR RNAs (crRNAs) for general orthopoxviruses and monkeypox virus-specific targets along with LbCas12a and fluorescent reporter. Recombinase polymerase amplification (RPA) of target loci was carried out followed by detection using the CRISPR-Cas12 CDM complex. Fluorescence-based read out can be monitored by a fluorescent reader and alternatively can also be visualized under a blue light illuminator by naked eye.ResultsMonkeypox-specific conserved sequences were identified inpolAgene which differ by a single nucleotide polymorphism (SNP) from all the viruses present in genus Orthopoxvirus. Our Cas12a-based assay was capable of specifically distinguishing monkeypox virus from other related orthopoxviruses with an LOD of 60 copies in 1 hour after the initiation of the reaction.ConclusionOur assay exhibits sensitive and specific detection of monkeypox virus which can prove to be of practical value for surveillance in areas infected with multiple orthopoxviruses, especially in hotspots of monkeypox virus infections.

show abstract

Section: Conclusion and Discussionmentioning

confidence: 99%

Section: Conclusion and Discussionmentioning

confidence: 99%

CRISPR/Cas12a assisted specific detection of monkeypox virus

Singh

Misra

Bindal

et al. 2022

Preprint

View full text Add to dashboard Cite

show abstract

“…According to the best of our acknowledge, there is only one short letter discussed the possibility of implementing CRISPR technology in MPXV detection, while details on pre-amplification, probe screening, analytical performance, and point-of-care testing (POCT) have not been included. 40 Herein, we proposed an ultrasensitive and rapid MPXV assay that integrates recombinase-aided amplification (RAA) with CRISPR/Cas12a (the RAA-Cas12a-MPXV assay) for the first time. After RAA pre-amplification of DNA templates (containing MPXV F3L gene), CRISPR/Cas12a-based amplicon recognition, and rapid cleavage of reporters, ultrasensitive detection of MPXV down to 10 1 copies/μL via fluorescence readout was accomplished, which is 1000-fold more sensitive than PCR.…”

Section: (Which Was Not Certified By Peer Review)mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

CRISPR/Cas12a-mediated ultrasensitive and on-site monkeypox viral testing

Zhao

Wang

et al. 2022

Preprint

View full text Add to dashboard Cite

The unexpected transmission of monkeypox virus (MPXV) from Central and West Africa to previously non-endemic locations is triggering a global panic. The ultrasensitive, rapid, and specific detection of MPXV is crucial for controlling its spreading, while such technology has rarely been reported. Herein, we proposed an MPXV assay combining recombinase-aided amplification (RAA) and CRISPR/Cas12a for the first time. This assay targeted MPXV F3L gene and yielded a low detection limit (LOD) of 101 copies/μL. Deriving from the high specificity nature of RAA and CRISPR/Cas12a, through rational optimizations of probes and conditions, this assay showed high selectivity that could distinguish MPXV from other orthopox viruses and current high-profile viruses. To facilitate on-site screening of potential MPXV carriers, a kit integrating lateral flow strips was developed, enabling naked-eye MPXV detection with a LOD of 104 copies/μL. Our RAA-Cas12a-MPXV assay was able to detect MPXV without the need for sophisticated operation and expensive equipment. We envision that this RAA-Cas12a-MPXV assay can be deployed in emerging viral outbreaks for on-site surveillance of MPXV.

show abstract

“…We read with great interest the paper by Yutong Sui et al, which developed a CRISPR-Cas12a-based detection assay for monkeypox virus (MPXV) 1 . Nucleic acid detection and antigen detection are two important methods for the diagnosis of the infection of the viruses, the two methods complement each other.…”

mentioning

confidence: 99%