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Updated: Nov 7, 2025
Detection of Invasive Pulmonary Aspergillosis in Haematological Malignancy Patients by using Lateral-flow Technology
Published on: March 22, 2012
Rapid Detection of Aspergillus fumigatus Using Multiple Cross Displacement Amplification Combined With
Luxi Jiang1, Xiaomeng Li1,2, Rumeng Gu1,2
1Department of Respiratory Medicine, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Hangzhou, China.
Abstract:
Aspergillus fumigatus is an opportunistic, ubiquitous, saprophytic mold which can cause infection in the lungs, nose, eyes, brain, and bones in humans, especially in immunocompromised patients. However, it is difficult to diagnose A. fumigatus infection quickly. Here, we introduce a new detection method, namely multiple cross displacement amplification (MCDA) combined with nanoparticle-based lateral flow biosensor (LFB) (MCDA-LFB), which was proved to be fast, reliable, and simple for detecting A. fumigatus. We designed a set of 10 primers targeting the gene annexin ANXC4 of A. fumigatus. The best MCDA condition is 66 °C for 35 min. The minimum concentration that can be detected by this method was 10 fg. In the case of 100 sputum samples, 20 (20%) and 15 (15%) samples were positive by MCDA-LFB and PCR method, respectively. MCDA-LFB and traditional culture method showed the same results. Compared with the culture method, the diagnostic accuracy of MCDA-LFB can reach 100%. It showed that the MCDA-LFB method has better detection ability than the PCR method. We found that the whole process could be controlled within 60 min including the preparation of DNA (20 min), MCDA reaction (35 min) and results reporting (2 min). These results show that this assay is suitable for the rapid, sensitive and specific detection of A. fumigatus in clinical samples.
Insights
A new multiple cross displacement amplification (MCDA) combined with nanoparticle-based lateral flow biosensor (LFB) method offers rapid, sensitive, and specific detection of Aspergillus fumigatus infections in clinical samples.
Area of Science:
- Medical Diagnostics
- Molecular Biology
- Mycology
Background:
- Aspergillus fumigatus is a common mold causing serious infections, particularly in immunocompromised individuals.
- Rapid and accurate diagnosis of A. fumigatus infections is challenging with current methods.
- Timely diagnosis is crucial for effective treatment and improved patient outcomes.
Purpose of the Study:
- To develop and validate a novel, rapid, and reliable detection method for Aspergillus fumigatus.
- To evaluate the diagnostic performance of the new method compared to existing techniques.
- To assess the feasibility of the method for clinical application.
Main Methods:
- Development of a multiple cross displacement amplification (MCDA) assay targeting the annexin ANXC4 gene of A. fumigatus.
- Integration of the MCDA assay with a nanoparticle-based lateral flow biosensor (LFB) for visual detection.
- Optimization of MCDA reaction conditions (66°C for 35 min) and determination of the limit of detection (10 fg).
- Validation using 100 clinical sputum samples and comparison with PCR and traditional culture methods.
Main Results:
- The MCDA-LFB method demonstrated high sensitivity, detecting down to 10 fg of target DNA.
- In sputum samples, MCDA-LFB identified 20% positive cases, compared to 15% by PCR.
- MCDA-LFB results were consistent with traditional culture methods, achieving 100% diagnostic accuracy.
- The entire detection process, including DNA preparation, can be completed within 60 minutes.
Conclusions:
- The MCDA-LFB assay is a fast, sensitive, and specific method for detecting Aspergillus fumigatus.
- This novel approach offers superior detection capabilities compared to the PCR method.
- The assay's speed and accuracy make it suitable for rapid clinical diagnosis of A. fumigatus infections.
