Related Experiment Video
Updated: Mar 26, 2026
Author Spotlight: Advancing Rapid Detection of Respiratory Pathogens Using Microfluidic Chip
Published on: March 29, 2024
Development and Evaluation of a Loop-Mediated Isothermal Amplification Method for Rapid Detection of Aspergillus
Qing Tang1, Shuguang Tian2, Nong Yu3
1Key Laboratory of Bio-resources and Eco-environment of the Ministry of Education, College of Life Sciences, Sichuan University, Chengdu, Sichuan, People's Republic of China.
Abstract:
Aspergillus fumigatusis a conditional pathogen and the major cause of life-threatening invasive aspergillosis (IA) in immunocompromised patients. The early and rapid detection ofA. fumigatusinfection is still a major challenge. In this study, the new member of the fungal annexin family, annexin C4, was chosen as the target to design a loop-mediated isothermal amplification (LAMP) assay for the rapid, specific, and sensitive detection ofA. fumigatus The evaluation of the specificity of the LAMP assay that was developed showed that no false-positive results were observed for the 22 non-A. fumigatusstrains, including 5 species of theAspergillusgenus. Its detection limit was approximately 10 copies per reaction in reference plasmids, with higher sensitivity than that of real-time quantitative PCR (qPCR) at 10(2)copies for the same target. Clinical samples from a total of 69 patients with probable IA (n =14) and possible IA (n= 55) were subjected to the LAMP assay, and positive results were found for the 14 patients with probable IA (100%) and 34 patients with possible IA (61.82%). When detection using the LAMP assay was compared with that using qPCR in the 69 clinical samples, the LAMP assay demonstrated a sensitivity of 89.19% and the concordance rate for the two methods was 72.46%. Accordingly, we report that a valuable LAMP assay for the rapid, specific, and simple detection ofA. fumigatusin clinical testing has been developed.
Insights
A new loop-mediated isothermal amplification (LAMP) assay effectively detects Aspergillus fumigatus, a common cause of invasive aspergillosis (IA). This rapid and sensitive method shows promise for early diagnosis in immunocompromised patients.
Area of Science:
- Mycology
- Molecular Biology
- Clinical Diagnostics
Background:
- Aspergillus fumigatus is a significant opportunistic pathogen causing life-threatening invasive aspergillosis (IA).
- Early and rapid detection of A. fumigatus infections remains a critical clinical challenge.
Purpose of the Study:
- To develop a loop-mediated isothermal amplification (LAMP) assay targeting the fungal annexin C4 gene for rapid, specific, and sensitive detection of A. fumigatus.
- To evaluate the performance of the developed LAMP assay in clinical samples.
Main Methods:
- Design and optimization of a LAMP assay targeting A. fumigatus annexin C4.
- Specificity testing against 22 non-A. fumigatus strains.
- Determination of the detection limit and comparison with real-time quantitative PCR (qPCR).
- Evaluation of the LAMP assay using 69 clinical samples from patients with suspected invasive aspergillosis.
Main Results:
- The LAMP assay demonstrated high specificity, with no cross-reactivity observed against related fungal species.
- The detection limit was approximately 10 copies per reaction, surpassing the sensitivity of qPCR (10^2 copies).
- In clinical samples, the LAMP assay correctly identified all 14 probable IA cases (100%) and 34 of 55 possible IA cases (61.82%).
- Compared to qPCR, the LAMP assay showed a sensitivity of 89.19% and a concordance rate of 72.46%.
Conclusions:
- A novel LAMP assay targeting annexin C4 provides a rapid, specific, and sensitive method for A. fumigatus detection.
- This LAMP assay holds significant potential for improving early diagnosis and management of invasive aspergillosis in clinical settings.
