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Related Concept Videos

The Antiviral System of Bacteria and Archaea: CRISPR01:23

The Antiviral System of Bacteria and Archaea: CRISPR

31
CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats is a adaptive immune system found in bacteria and archaea that protects against viral infections. This system enables prokaryotic cells to identify, remember, and neutralize foreign genetic elements, primarily bacteriophages, by storing fragments of the invader’s DNA as a genetic memory.The CRISPR immune response begins during an initial infection. Cas (CRISPR-associated) proteins play a central role in this...
31

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Related Experiment Video

Updated: Jul 12, 2025

Rapid Diagnosis of Avian Influenza Virus in Wild Birds: Use of a Portable rRT-PCR and Freeze-dried Reagents in the Field
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CRISPR-Cas13a-based detection method for avian influenza virus.

Yuhan Wu1, Jiaxing Zhan1, Zhaomeng Shan1

  • 1Key Laboratory of Livestock Infectious Diseases, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang, China.

Frontiers in Microbiology
|October 27, 2023
PubMed
Summary

A new CRISPR-Cas13a diagnostic platform rapidly detects avian influenza virus (AIV) subtypes H1-H16. This portable assay simplifies detection without nucleic acid extraction, offering potential for point-of-care testing in avian disease control.

Keywords:
CRISPR-cas13aavian influenza viruslateral flow dipstick assayrecombinase polymerase amplificationuniversal detection

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Area of Science:

  • Veterinary Virology
  • Molecular Diagnostics
  • Biosensing Technologies

Background:

  • Avian influenza virus (AIV) poses significant economic and public health risks.
  • Rapid, sensitive, and portable diagnostic tools are crucial for effective AIV surveillance and control.
  • Current diagnostic methods can be time-consuming and require specialized laboratory equipment.

Purpose of the Study:

  • To develop a novel clustered regularly interspaced short palindromic repeats (CRISPR)-Cas13a based platform for sensitive and specific AIV detection.
  • To enable rapid, portable, and simplified detection of AIV, suitable for point-of-care applications.
  • To validate the assay's performance against established diagnostic techniques and clinical samples.

Main Methods:

  • Development of a CRISPR-Cas13a system for specific AIV detection across H1-H16 subtypes.
  • Integration of fluorescence and lateral flow readouts for versatile detection.
  • Implementation of a one-tube method combining reverse transcription, recombinase polymerase amplification, and CRISPR-Cas13a detection.
  • Introduction of a sample heating step to eliminate the need for nucleic acid extraction.

Main Results:

  • The developed assay demonstrated high specificity for AIV, with no cross-reactivity against other common avian viruses.
  • The limit of detection was as low as 69 copies/μL (fluorescence) and 690 copies/μL (lateral flow).
  • The assay achieved 100% consistency with quantitative real-time polymerase chain reaction (qRT-PCR) in clinical sample analysis.
  • A simplified workflow allowing detection from unextracted samples was successfully implemented.

Conclusions:

  • The CRISPR-Cas13a platform offers a rapid, sensitive, and specific method for AIV detection.
  • The simplified, one-tube, no-extraction protocol reduces turnaround time and contamination risk.
  • This technology holds significant promise for point-of-care diagnostics in avian influenza surveillance and management.