Related Experiment Video
Updated: May 11, 2026
Antigen-Capture Enzyme-Linked Immunosorbent Assay for Specific Detection of Mycoplasma pneumoniae
Published on: February 24, 2023
CHAMP: A Centrifugal Microfluidics-Based CRISPR/Cas12b-Combined Real-Time LAMP One-Pot Method for Mycoplasma
Wenwen Jing1, Tong Zhang2, Xiangyang Min3
1Department of Medical Microbiology and Parasitology, School of Basic Medical Sciences, Fudan University, Shanghai 200032, P. R. China.
Abstract:
The Mycoplasma pneumoniae outbreak poses health risks to community residents. However, it still has limitations for current clinical diagnostic methods (qPCR nucleic acid assay or IgM immunoassay), including specialized handling, expensive equipment, prolonged turnaround time, and false positives and negatives, highlighting the need to improve clinical diagnostic methods. Herein, we present a novel centrifugal microfluidics-based method for rapidly diagnosing M. pneumoniae infections (CHAMP system). This user-friendly method combines CRISPR/Cas12b and real-time loop-mediated isothermal amplification (LAMP) in a one-pot reaction, offering high sensitivity, specificity, and simplicity for methodology. By adding fully automated nucleic acid magnetic bead-extracted samples to a prepackaged centrifugal microfluidics chip, 48 samples can be automated tested simultaneously within 15 to 60 min at 60 °C. 427 clinical nasopharyngeal swab specimens were used for validation, demonstrating good positive and negative predictive values and good diagnostic sensitivity, specificity, and significant time savings. This method is particularly suitable for detecting low nucleic acid copies of M. pneumoniae samples.
Insights
A new CHAMP system rapidly diagnoses Mycoplasma pneumoniae infections using CRISPR/Cas12b and LAMP technology. This user-friendly method offers a faster, more accurate clinical diagnostic alternative for M. pneumoniae detection.
Area of Science:
- Medical Diagnostics
- Molecular Biology
- Biotechnology
Background:
- Mycoplasma pneumoniae outbreaks present significant public health risks.
- Current diagnostic methods like qPCR and IgM immunoassays have limitations, including cost, time, and accuracy.
- There is a critical need for improved, rapid, and accessible clinical diagnostic tools.
Purpose of the Study:
- To develop and validate a novel, user-friendly centrifugal microfluidics-based diagnostic system for Mycoplasma pneumoniae infections.
- To enhance diagnostic speed, sensitivity, and specificity compared to existing methods.
- To provide a simplified workflow for clinical settings.
Main Methods:
- Development of the CHAMP system, integrating CRISPR/Cas12b with real-time loop-mediated isothermal amplification (LAMP) in a one-pot reaction.
- Utilizing a prepackaged centrifugal microfluidics chip for automated sample processing and testing.
- Validation using 427 clinical nasopharyngeal swab specimens.
Main Results:
- The CHAMP system demonstrated high sensitivity, specificity, and simplicity.
- Simultaneous automated testing of 48 samples was achieved within 15 to 60 minutes at 60 °C.
- The method showed good positive and negative predictive values and significant time savings.
- Effective detection of low nucleic acid copies of Mycoplasma pneumoniae was confirmed.
Conclusions:
- The novel CHAMP system offers a rapid, sensitive, and specific diagnostic solution for Mycoplasma pneumoniae infections.
- This user-friendly, microfluidics-based approach addresses limitations of current clinical diagnostic methods.
- The CHAMP system is well-suited for widespread clinical application, especially for detecting low pathogen loads.
