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Updated: May 1, 2026
Detection of Invasive Pulmonary Aspergillosis in Haematological Malignancy Patients by using Lateral-flow Technology
Published on: March 22, 2012
Development and validation of an HPLC-MS/MS method for the early diagnosis of aspergillosis
Letícia B Cerqueira1, Thais M G de Francisco1, João C Gasparetto1
1Department of Pharmacy, Federal University of Paraná, Curitiba, Paraná, Brazil.
Abstract:
Invasive aspergillosis is an opportunistic infection that is mainly caused by Aspergillus fumigatus, which is known to produce several secondary metabolites, including gliotoxin, the most abundant metabolite produced during hyphal growth. The diagnosis of invasive aspergillosis is often made late in the infection because of the lack of reliable and feasible diagnostic techniques; therefore, early detection is critical to begin treatment and avoid more serious complications. The present work reports the development and validation of an HPLC-MS/MS method for the detection of gliotoxin in the serum of patients with suspected aspergillosis. Chromatographic separation was achieved using an XBridge C18 column (150 × 2.1 mm id; 5 mm particle size) maintained at 25 °C with the corresponding guard column (XBridge C18, 10 × 2.1 mm id, 5 mm particle size). The mobile phase was composed of a gradient of water and acetonitrile/water (95:5 v/v), both containing 1 mM ammonium formate with a flow rate of 0.45 mL min(-1). Data from the validation studies demonstrate that this new method is highly sensitive, selective, linear, precise, accurate and free from matrix interference. The developed method was successfully applied to samples from patients suspected of having aspergillosis. Therefore, the developed method has considerable potential as a diagnostic technique for aspergillosis.
Insights
A new HPLC-MS/MS method detects gliotoxin in patient serum, enabling earlier diagnosis of invasive aspergillosis. This advancement is crucial for timely treatment and improved patient outcomes in fungal infections.
Area of Science:
- Clinical Chemistry
- Analytical Chemistry
- Mycology
Background:
- Invasive aspergillosis, often caused by Aspergillus fumigatus, is difficult to diagnose early due to limited reliable methods.
- Aspergillus fumigatus produces gliotoxin, a key metabolite during hyphal growth, making it a potential biomarker.
- Late diagnosis of invasive aspergillosis leads to severe complications and poorer prognoses.
Purpose of the Study:
- To develop and validate a highly sensitive and specific HPLC-MS/MS method for detecting gliotoxin in human serum.
- To establish a feasible diagnostic tool for the early identification of invasive aspergillosis.
Main Methods:
- Development of an HPLC-MS/MS method utilizing an XBridge C18 column and a gradient mobile phase.
- Validation of the method for sensitivity, selectivity, linearity, precision, accuracy, and matrix interference.
- Application of the validated method to serum samples from patients with suspected aspergillosis.
Main Results:
- The developed HPLC-MS/MS method demonstrated high sensitivity, selectivity, linearity, precision, and accuracy.
- The method proved to be free from matrix interference in patient serum samples.
- Successful application of the method to clinical samples from patients with suspected invasive aspergillosis.
Conclusions:
- The developed HPLC-MS/MS method is a reliable and sensitive technique for gliotoxin detection.
- This method shows significant potential as an early diagnostic tool for invasive aspergillosis.
- Early detection via gliotoxin measurement can facilitate prompt treatment and improve patient outcomes.
