Related Experiment Video
Updated: Jul 8, 2026
Detection of Invasive Pulmonary Aspergillosis in Haematological Malignancy Patients by using Lateral-flow Technology
Published on: March 22, 2012
Establishing a pulmonary aspergillus fumigatus infection diagnostic platform based on RPA-CRISPR-Cas12a
Chunhui Lin1,2, Jing Zhou1,2, Nana Gao1,2
1Department of Clinical Laboratory, the First Affiliated Hospital of Anhui Medical University, Hefei, People's Republic of China.
Abstract:
In this study, we devised a diagnostic platform harnessing a combination of recombinase polymerase amplification (RPA) and the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a system. Notably, this platform obviates the need for intricate equipment and finds utility in diverse settings. Two result display methods were incorporated in this investigation: the RPA-Cas12a-fluorescence method and the RPA-Cas12a-LFS (lateral flow strip). Upon validation, both display platforms exhibited no instances of cross-reactivity, with seven additional types of fungal pathogens responsible for respiratory infections. The established detection limit was ascertained to be as low as 102 copies/µL. In comparison to fluorescence quantitative PCR, the platform demonstrated a sensitivity of 96.7%, a specificity of 100%, and a consistency rate of 98.0%.This platform provides expeditious, precise, and on-site detection capabilities, thereby rendering it a pivotal diagnostic instrument amenable for deployment in primary healthcare facilities and point-of-care settings.
Insights
This study presents a novel diagnostic platform using recombinase polymerase amplification (RPA) and CRISPR/Cas12a for rapid, on-site detection of respiratory fungal pathogens. It offers high sensitivity and specificity without complex equipment.
Area of Science:
- Molecular Biology
- Biotechnology
- Diagnostic Technologies
Background:
- Respiratory fungal infections pose a significant health burden.
- Current diagnostic methods often require specialized equipment and trained personnel.
- There is a need for rapid, sensitive, and accessible diagnostic tools.
Purpose of the Study:
- To develop and validate a novel diagnostic platform for detecting respiratory fungal pathogens.
- To integrate recombinase polymerase amplification (RPA) with the CRISPR/Cas12a system for enhanced detection.
- To evaluate the platform's performance using both fluorescence and lateral flow strip (LFS) detection methods.
Main Methods:
- Combined RPA and CRISPR/Cas12a for nucleic acid detection.
- Utilized two readout formats: RPA-Cas12a-fluorescence and RPA-Cas12a-LFS.
- Tested for cross-reactivity against seven common fungal pathogens.
- Determined the limit of detection (LOD) and compared performance against quantitative PCR (qPCR).
Main Results:
- The platform demonstrated high specificity, with no cross-reactivity against other fungal pathogens.
- Achieved a low limit of detection (LOD) of 10^2 copies/µL.
- Exhibited high sensitivity (96.7%), specificity (100%), and consistency (98.0%) compared to qPCR.
- Both fluorescence and LFS methods provided reliable results.
Conclusions:
- The developed RPA-CRISPR/Cas12a platform offers a rapid, precise, and equipment-light diagnostic solution.
- The platform is suitable for point-of-care and primary healthcare settings.
- This technology has the potential to significantly improve the diagnosis and management of respiratory fungal infections.
More Related Videos
10:16Author Spotlight: Development of Simplified CRISPR-Based Tests for Rapid Detection of Infectious Diseases
Published on: August 16, 2024
07:59Rapid and Specific Detection of Acinetobacter baumannii Infections Using a Recombinase Polymerase Amplification/Cas12a-based System
Published on: April 25, 2025
