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Updated: Jul 15, 2026
Detection of Invasive Pulmonary Aspergillosis in Haematological Malignancy Patients by using Lateral-flow Technology
Published on: March 22, 2012
Detection of Aspergillus spp. in biological samples by real-time PCR
Radka Bolehovska1, Lenka Pliskova, Vladimir Buchta
1Department of Clinical Biochemistry and Diagnostics, Faculty of Medicine and University Hospital, Charles University, Hradec Kralove, 500 05, Czech Republic. bolehrad@fnhk.cz
Background:
Recently, the proportion of invasive infections caused by the filamentous fungi of the Aspergillus genus are growing in immunocompromised persons particularly in transplant recipients and neutropenic patients. Unfortunately, laboratory diagnostics of invasive aspergillosis remains extremely difficult, mainly with regard to the sensitivity of the methods and to the correct interpretation of the results in particular.
Aim:
The aim of this work was to design a standard and reproducible Aspergillus DNA detection method and its validation. The second aim was to practically use this method for diagnosis of Aspergillus DNA in various samples in patients.
Method:
Real-time PCR with two hybridization probes. Amplification and on-line quantification was carried out on a LightCycler 1.5 Instrument.
Results:
Specificity of the reaction was tested for A. fumigatus, A. flavus, A. niger and A. terreus, and its sensitivity was determined at 5 copies per ml. The reproducibility of the results was comparable to other methods, reported in the literature. Applicability of the real-time PCR was assessed for detection of Aspergillus DNA in 354 various clinical samples taken from 179 patients at risk of invasive aspergillosis over the period of 33 months. Of 354 samples 103 (29.10 %) taken from 65 patients (36.31 %) were evaluated as positive. Over one year, the percentage of positive samples was mostly about 30 % or less per month.
Conclusions:
Our results demonstrate the high sensitivity, specificity and reproducibility of this technique, and its usefulness for rapid laboratory diagnosis of invasive aspergillosis.
Insights
A new real-time PCR method accurately detects Aspergillus DNA in patients, improving diagnosis of invasive aspergillosis. This sensitive and specific technique aids in early identification of fungal infections in immunocompromised individuals.
Area of Science:
- Medical Mycology
- Molecular Diagnostics
- Infectious Diseases
Background:
- Invasive infections caused by Aspergillus are increasing in immunocompromised patients.
- Current laboratory diagnostics for invasive aspergillosis are challenging due to sensitivity and interpretation issues.
Purpose of the Study:
- To develop and validate a standardized, reproducible method for Aspergillus DNA detection.
- To apply this method for diagnosing Aspergillus DNA in patient samples.
Main Methods:
- Real-time PCR with two hybridization probes.
- Amplification and online quantification performed on a LightCycler 1.5 Instrument.
Main Results:
- The method demonstrated high specificity for Aspergillus species and a sensitivity of 5 copies/mL.
- Tested on 354 clinical samples from 179 patients, 29.10% of samples and 36.31% of patients were positive for Aspergillus DNA.
- Reproducibility was comparable to existing methods.
Conclusions:
- The developed real-time PCR technique is highly sensitive, specific, and reproducible.
- This method is useful for the rapid laboratory diagnosis of invasive aspergillosis.
