Visual detection of human metapneumovirus using CRISPR‐Cas12a diagnostics

Qian, Weidong; Huang, Jie; Wang, Ting; He, Xiaoxian; Xu, Guozhang

doi:10.1016/j.virusres.2021.198568

Cited by 10 publications

(11 citation statements)

References 28 publications

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“…This method for detection of infectious agents has been developed and widely evaluated. Compared with routine PCR methods, CRISPR has several advantages, such as a short turn round time, low cost, easy-to-use, and no requirement of expensive equipment [ 16 , 31 , 32 ]. For example, CRISPR-Cas12a technology was evaluated in the diagnosis of human papillomavirus, and the data suggested that this tool could achieve results within 35 min with a low detection limit [ 33 ].…”

Section: Discussionmentioning

confidence: 99%

“…Recently, the clustered regularly interspaced short palindromic repeats (CRISPR) system has been introduced in the diagnosis of infectious diseases, such as tuberculosis [ 15 ], metapneumovirus [ 16 ], and malaria [ 17 ]. Such assays employ the trans cleavage activity of a Cas12/gRNA complex bound to an amplicon recognition sequence to cleave and derepress a quenched fluorescent oligonucleotide reporter that is present at a high concentration.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Evaluation of an automated CRISPR-based diagnostic tool for rapid detection of COVID-19

Xu¹,

Ma²,

Song³

et al. 2023

Heliyon

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Evaluation of an automated CRISPR-based diagnostic tool for rapid detection of COVID-19

Xu¹,

Ma²,

Song³

et al. 2023

Heliyon

View full text Add to dashboard Cite

“…The minimum detection limit was 9.65×10 2 copies/ml, and the detection coincidence rate with qRT-PCR was 98.3%( Qian et al., 2021b ). RPA-CRISPR/Cas12a could also detect Human metapneumovirus ( Qian et al., 2021a ). CRISPR/lwCas13a was another protein that could be used for nucleic acid detection, it had been successfully combined with RPA to detect the African swine fever virus (ASFV) ( Ren et al., 2021a ) and rabies virus ( Ren et al., 2021b ).…”

Section: The Application Of Rpamentioning

confidence: 99%

Recent advances in recombinase polymerase amplification: Principle, advantages, disadvantages and applications

Tan

Liao

Liu

et al. 2022

Front. Cell. Infect. Microbiol.

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After the outbreak of SARS-CoV-2, nucleic acid testing quickly entered people’s lives. In addition to the polymerase chain reaction (PCR) which was commonly used in nucleic acid testing, isothermal amplification methods were also important nucleic acid testing methods. Among several common isothermal amplification methods like displaced amplification, rolling circle amplification, and so on, recombinase polymerase amplification (RPA) was recently paid more attention to. It had the advantages like a simple operation, fast amplification speed, and reaction at 37-42°C, et al. So it was very suitable for field detection. However, there were still some disadvantages to RPA. Herein, our review mainly summarized the principle, advantages, and disadvantages of RPA. The specific applications of RPA in bacterial detection, fungi detection, virus detection, parasite detection, drug resistance gene detection, genetically modified food detection, and SARS-CoV-2 detection were also described. It was hoped that the latest research progress on RPA could be better delivered to the readers who were interested in RPA.

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“…Additionally, cleavage results of CRISPR/Cas12a system can be visualized using lateral flow biosensor (LFB) or fluorescence reader by introducing ssDNA reporter labeled with fluorophore and biotin or alternatively with fluorophore and quencher into an in vitro reaction (Xiong et al, 2020;Zhu et al, 2021). Now, CRISPR/Cas12a detection integrated with RPA has been successfully applied in rapid and accurate detection of various pathogens including viral pathogens (such as SARS-CoV-2, human metapneumovirus, human papillomavirus and African swine fever; Bai et al, 2019;Yin et al, 2020;Qian et al, 2021;Sun et al, 2021;Talwar et al, 2021), fungal pathogens (for example Elsinoë fawcettii; Shin et al, 2021), Bacterial pathogens (such as Escherichia coli, Listeria monocytogenes, Staphylococcus aureus, Vibrio parahaemolyticus, Salmonella sp., Xanthomonas arboricola and Pseudomonas aeruginosa; Cai et al, 2021;Liu et al, 2021;Luo et al, 2021). In this study, RPA coupled with CRISPR/Cas12a (RPA-CRISPR/Cas12a) has been developed for MP detection.…”

Section: Introductionmentioning

confidence: 99%

Development of a Rapid and Efficient RPA-CRISPR/Cas12a Assay for Mycoplasma pneumoniae Detection

Xiao

Yang

et al. 2022

Front. Microbiol.

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Mycoplasma pneumoniae (MP) is a one of most common pathogen in causing respiratory infection in children and adolescents. Rapid and efficient diagnostic methods are crucial for control and treatment of MP infections. Herein, we present an operationally simple, rapid and efficient molecular method for MP identification, which eliminates expensive instruments and specialized personnel. The method combines recombinase polymerase amplification (RPA) with clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated proteins (Cas) 12a-based detection, with an optimal procedure less than 1 h from sample to result including DNA extraction (25 min), RPA reaction (39°C for 15-20 min), CRISPR/Cas12a detection (37°C for 10 min) and visual detection by naked eyes (2 min). This diagnostic method shows high sensitivity (two copies per reaction) and no cross-reactivity against other common pathogenic bacteria. Preliminary evaluation using 201 clinical samples shows sensitivity of 99.1% (107/108), specificity of 100% (93/93) and consistency of 99.5% (200/201), compared with real-time PCR method. The above data demonstrate that our developed method is reliable for rapid diagnosis of MP. In conclusion, the RPA-CRISPR/Cas12a has a great potential to be as a useful tool for reliable and quick diagnosis of MP infection, especially in primary hospitals with limited conditions.

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Visual detection of human metapneumovirus using CRISPR‐Cas12a diagnostics

Cited by 10 publications

References 28 publications

Evaluation of an automated CRISPR-based diagnostic tool for rapid detection of COVID-19

Evaluation of an automated CRISPR-based diagnostic tool for rapid detection of COVID-19

Recent advances in recombinase polymerase amplification: Principle, advantages, disadvantages and applications

Development of a Rapid and Efficient RPA-CRISPR/Cas12a Assay for Mycoplasma pneumoniae Detection

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