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Updated: Aug 6, 2026
Optimized PCR-based Detection of Mycoplasma
Published on: June 20, 2011
EasyNAT MP Assay: A Simple, Rapid, and Low-Cost Method to Detect Mycoplasma pneumoniae Using Cross-Priming
Guang Chen1, Junwei Yu2, Hanlu Chen3
1Department of Clinical Laboratory, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, 310012, Zhejiang, People's Republic of China.
Background:
Mycoplasma pneumoniae (MP) is the most common pathogen of atypical pneumonia and the main cause of community-acquired pneumonia (CAP) in infants and older adults. This study aimed at investigating a method based on the cross-priming amplification (CPA) technique for the rapid detection of MP in clinical specimens collected from patients with CAP.
Methods:
The sensitivity and specificity of the EasyNAT MP assay were determined. Oropharyngeal swab specimens were collected from 162 in-patients of Hangzhou First People's Hospitals from January 2018 to December 2020. The patients were aged between 1 and 15 years with symptoms, signs, and chest radiographs consistent with CAP. This study evaluated the presence of MP in the clinical specimens using the EasyNAT method and the conventional fluorescence quantitative PCR technique.
Results:
The limit of detection using the EasyNAT MP assay was 500 copies/mL, while the test results of the other 13 common pathogens causing CAP or colonizing in the upper respiratory tract showed no cross-reactivity. Of 162 specimens, EasyNAT MP gave a positive indication in 82 specimens. Compared with conventional fluorescence quantitative PCR, the positive coincidence rate and the negative coincidence rate of EasyNAT MP was found to be 100.00% and 97.56%, respectively. Of the 82 specimens, two specimens were determined to be negative by the conventional fluorescence quantitative PCR, but were positive for EasyNAT MP. The two samples were re-extracted and confirmed to be positive by conventional fluorescence quantitative PCR.
Conclusion:
EasyNAT MP is suitable as an initial test for MP diagnosis due to its simplicity, low turnaround time, and high sensitivity and specificity.
Insights
This study validates the EasyNAT MP assay for rapid Mycoplasma pneumoniae detection in children with community-acquired pneumonia (CAP). The assay shows high sensitivity and specificity, making it a reliable diagnostic tool.
Area of Science:
- Microbiology
- Infectious Diseases
- Diagnostic Technologies
Background:
- Mycoplasma pneumoniae (MP) is a primary cause of atypical pneumonia and community-acquired pneumonia (CAP) in pediatric populations.
- Accurate and rapid diagnosis of MP is crucial for effective patient management.
Purpose of the Study:
- To evaluate the performance of the EasyNAT MP assay, a novel method based on cross-priming amplification (CPA), for the rapid detection of MP in clinical specimens from pediatric CAP patients.
- To determine the sensitivity and specificity of the EasyNAT MP assay in comparison to conventional fluorescence quantitative PCR.
Main Methods:
- Oropharyngeal swab specimens were collected from 162 pediatric patients diagnosed with CAP.
- The EasyNAT MP assay was used to detect MP, and results were compared with conventional fluorescence quantitative PCR.
- The limit of detection and cross-reactivity with other respiratory pathogens were assessed for the EasyNAT MP assay.
Main Results:
- The EasyNAT MP assay demonstrated a limit of detection of 500 copies/mL with no cross-reactivity against 13 other common respiratory pathogens.
- Out of 162 specimens, 82 tested positive using the EasyNAT MP assay.
- The assay achieved 100.00% positive and 97.56% negative coincidence rates compared to conventional fluorescence quantitative PCR, with discrepancies resolved by re-testing.
Conclusions:
- The EasyNAT MP assay is a simple, rapid, and highly sensitive and specific diagnostic tool for MP.
- Its performance characteristics make it suitable as an initial diagnostic test for MP in pediatric CAP cases.
