ArticleBiotechnology and bioengineering2021
A CRISPR/Cas9 eraser strategy for contamination-free PCR end-point detection.
Article in Biotechnology and bioengineering, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed, 34 citations in OpenAlex.
- A One-Pot CRISPR/Cas12a-Based Platform for Contamination-Free Nucleic Acid Amplification Detection.Biosensors · 2026Article
- Effects of Phytosterols on Growth Performance, Serum Indexes, and Fecal Microbiota in Finishing Pigs.Animals : an open access journal from MDPI · 2025Article
- CRISPR/Cas system and its application in the diagnosis of animal infectious diseases.FASEB journal : official publication of the Federation of American Societies for Experimental Biology · 2024Review
- Effects of maternal hawthorn-leaf flavonoid supplementation on the intestinal development of offspring chicks.Poultry science · 2024Article
- CRISPR-Cas9-mediated host signal reduction for 18S metabarcoding of host-associated eukaryotes.Molecular ecology resources · 2024Article
- Progress and Perspective of CRISPR-Cas9 Technology in Translational Medicine.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2023Review
- Carryover Contamination-Controlled Amplicon Sequencing Workflow for Accurate Qualitative and Quantitative Detection of Pathogens: a Case Study on SARS-CoV-2.Microbiology spectrum · 2023Article
- Paper microfluidics with deep learning for portable intelligent nucleic acid amplification tests.Talanta · 2023Article
- Review
- Non-nucleic acid extraction and ultra-sensitive detection of African swine fever virus via CRISPR/Cas12a.Applied microbiology and biotechnology · 2022Article
- Improving the specificity of nucleic acid detection with endonuclease-actuated degradation.Communications biology · 2022Article
- Design and storage stability of reference materials for microfluidic quantitative PCR-based equine gene doping tests.Journal of equine science · 2021Article
- CRISPR-Cas based virus detection: Recent advances and perspectives.Biosensors & bioelectronics · 2021Review
- Potential of CRISPR/Cas system in the diagnosis of COVID-19 infection.Expert review of molecular diagnostics · 2021Review
- A CRISPR/Cas9 eraser strategy for contamination-free PCR end-point detection.Biotechnology and bioengineering · 2021Article
Corrections and comments
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Authors and funding
6 authors at 2 institutions in 1 country.
Funding
Abstract
Polymerase chain reaction (PCR), a central technology for molecular diagnostics, is highly sensitive but susceptible to the risk of false positives caused by aerosol contamination, especially when an end-point detection mode is applied. Here, we proposed a solution by designing a clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 eraser strategy for eliminating potential contamination amplification. The CRISPR/Cas9 engineered eraser is firstly adopted into artpcr reverse-transcription PCR (RT-PCR) system to achieve contamination-free RNA detection. Subsequently, we extended this CRISPR/Cas9 eraser to the PCR system. We engineered conventional PCR primers to enable the amplified products to contain an implanted NGG (protospacer adjacent motif, PAM) site, which is used as a code for specific CRISPR/Cas9 recognition. Pre-incubation of Cas9/sgRNA with PCR mix leads to a selective cleavage of contamination amplicons, thus only the template DNA is amplified. The developed CRISPR/Cas9 eraser, adopted by both RT-PCR and PCR systems, showed high-fidelity detection of SARS-CoV-2 and African swine fever virus with a convenient strip test.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.