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Updated: Jun 10, 2026
Detection of Invasive Pulmonary Aspergillosis in Haematological Malignancy Patients by using Lateral-flow Technology
Published on: March 22, 2012
Primary diagnostic approaches of invasive aspergillosis--molecular testing
1Groupe hospitalier Chenevier-Mondor, Hôpital Henri Mondor-APHP,Laboratoire de Parasitologie-Mycologie, Créteil, France. bretagne@univ-paris12.fr
Abstract:
The PCR methods published for the diagnosis of invasive aspergillosis (IA) are diverse in terms of amplification protocols and methods, equipment, fluorescent detection dyes, PCR chemistries, and clinical specimens used. This explains why PCR is still not included in the revised EORTC/MSG definitions of IA despite encouraging results. Therefore, achieving consensual PCR procedures at the international level is mandatory. When using PCR as a diagnostic tool, emphasis must be put on limiting false positive results due to contamination either with previously amplified products or with environmental commensals. Internal amplification controls are compulsory to evidence false negative results. For most of these aspects, quantitative PCR (qPCR) should improve both the results' reliability and the clinicians' confidence. A checklist of items (Minimum information for publication of quantitative real-time PCR experiments) has been proposed to help scientists and reviewers. Currently, the main limitation relies in the DNA extraction procedure the choice of which dramatically depends on the still unknown origin of the Aspergillus DNA to amplify. There is an urgent need for basic studies to elucidate the origin and kinetics of Aspergillus DNA in blood. Once a technical consensus is achieved, clinical studies should be initiated to integrate qPCR in the diagnostic armentarium of IA.
Insights
Standardizing PCR methods is crucial for diagnosing invasive aspergillosis (IA). Quantitative PCR (qPCR) offers improved reliability, but DNA extraction challenges and a need for clinical validation remain. Further research is essential for IA diagnosis.
Area of Science:
- Medical Mycology
- Molecular Diagnostics
- Infectious Diseases
Background:
- Published PCR methods for invasive aspergillosis (IA) diagnosis lack standardization.
- This heterogeneity hinders PCR's inclusion in diagnostic criteria despite promising results.
- International consensus on PCR procedures is necessary for reliable IA diagnosis.
Purpose of the Study:
- To highlight the need for standardized PCR protocols in invasive aspergillosis diagnosis.
- To emphasize the potential of quantitative PCR (qPCR) in improving diagnostic accuracy.
- To identify current limitations and future directions for PCR-based IA diagnostics.
Main Methods:
- Review of diverse PCR amplification protocols, detection methods, and clinical specimens used for IA.
- Discussion on the importance of internal controls to prevent false negatives and minimize false positives.
- Introduction of Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines.
Main Results:
- Significant variability exists in PCR techniques, impacting diagnostic reliability.
- Quantitative PCR (qPCR) shows potential for enhanced result validity and clinical confidence.
- DNA extraction remains a major hurdle, dependent on the unknown origin of Aspergillus DNA.
Conclusions:
- Standardized international PCR procedures are mandatory for reliable invasive aspergillosis diagnosis.
- Further research is urgently needed to understand Aspergillus DNA origin and kinetics in blood.
- Clinical studies are required to integrate validated qPCR into the diagnostic armamentarium for IA.
