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Related Concept Videos

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CRISPR

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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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CRISPR and crRNAs02:53

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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
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Updated: Jun 4, 2025

Field-Deployable Candidatus Liberibacter asiaticus Detection Using Recombinase Polymerase Amplification Combined with CRISPR-Cas12a
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CRISPR-Cas12a2-based rapid and sensitive detection system for target nucleic acid.

Helin Yu1, Meng Feng2, Chuncao Liu1

  • 1Shanghai Institute of Infectious Disease and Biosecurity, Fudan University, Shanghai 200032, China; Department of Medical Microbiology and Parasitology, School of Basic Medical Sciences, Fudan University, Shanghai 20032, China.

International Journal of Biological Macromolecules
|December 20, 2024
PubMed
Summary

This study introduces novel PCR-SuCas12a2 and RPA-SuCas12a2 detection methods for infectious diseases. PCR-SuCas12a2 demonstrates ultra-sensitivity and high specificity, outperforming RPA-SuCas12a2 in detecting pathogens like Entamoeba histolytica.

Keywords:
CRISPR-SuCas12a2Fluorescence quantitationISOthermal reactionNucleic acid detectionTarget RNA

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DNA Virus Detection System Based on RPA-CRISPR/Cas12a-SPM and Deep Learning

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Public Health

Background:

  • Infectious diseases pose significant public health challenges, necessitating rapid and effective diagnostic tools.
  • Current detection methods often require complex laboratory setups and can be time-consuming.

Purpose of the Study:

  • To develop novel, sensitive, and specific nucleic acid detection platforms using a novel RNA-targeting nuclease, SuCas12a2.
  • To evaluate the performance of these platforms, named PCR-SuCas12a2 and RPA-SuCas12a2, for detecting specific pathogens.

Main Methods:

  • Coupling SuCas12a2 with Polymerase Chain Reaction (PCR) and Recombinase Polymerase Amplification (RPA) to create PCR-SuCas12a2 and RPA-SuCas12a2 assays.
  • Utilizing SuCas12a2's collateral cleavage activity on single-stranded RNA (ssRNA) probes, triggered by target-specific binding to CRISPR RNA (crRNA), to generate fluorescent signals.
  • Incorporating a T7 promoter for simultaneous transcription and detection in a single tube.
  • Validating the assays using Entamoeba histolytica (E. histolytica) and Mycoplasma pneumoniae (M. pneumoniae) as model organisms and clinical samples.

Main Results:

  • PCR-SuCas12a2 achieved ultra-high sensitivity, detecting as low as one copy per reaction for E. histolytica and M. pneumoniae, with no cross-reactivity.
  • RPA-SuCas12a2 demonstrated a sensitivity of 10^2 copies per reaction, which was less sensitive than the PCR-SuCas12a2 method.
  • Both assays exhibited 100% specificity when tested with clinical samples from suspected E. histolytica and M. pneumoniae infections.

Conclusions:

  • The developed PCR-SuCas12a2 assay offers excellent sensitivity and specificity for pathogen detection.
  • The SuCas12a2-based detection platforms show significant potential for rapid, point-of-care testing (POCT) applications in infectious disease diagnostics.
  • These novel methods could aid in the effective management and control of infectious disease outbreaks.