Identification of Aspergillus Conidia in Bronchoalveolar Lavage Using Offline Combination of Capillary Electrophoresis in Supercritical Water-Treated Fused Silica Capillary and Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

• Published in: Analytical chemistry (Washington) 2020-06, Vol.92 (11), p.7588-7595
• Main Authors: Šalplachta, Jiří, Horká, Marie, Karásek, Pavel, Růžička, Filip, Roth, Michal
• Format: Article
• Language: English
• Subjects: Alveoli; Analytical chemistry; Aspergillus; Aspergillus - isolation & purification; Bronchoalveolar Lavage; Bronchoalveolar Lavage Fluid - microbiology; Bronchoscopy; Bronchus; Capillary electrophoresis; Chemistry; Conidia; Desorption; Diagnosis; Electrokinetics; Electrophoresis; Electrophoresis, Capillary; Etching; Fused silica; Humans; Ionization; Ions; Isotachophoresis; Lavage; Mass spectrometry; Mass spectroscopy; Parenchyma; Scientific imaging; Silica; Silicon dioxide; Silicon Dioxide - chemistry; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Spectroscopy; Water - chemistry; Water treatment
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/acs.analchem.0c00165

Diagnosis of fungal infection in lung parenchyma is relatively difficult. Bronchoscopy with bronchoalveolar lavage is very useful in its diagnosing. Therefore, a method for rapid online concentration and analysis of conidia in bronchoalveolar lavage fluid using the combination of transient isotachophoresis (tITP) and micellar electrokinetic chromatography (MEKC) with subsequent off-line identification of the separated conidia by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is described in this study. In the proposed procedure, conidia were first dynamically adhered onto the roughened part of the inner surface of a fused silica capillary prepared by etching with supercritical water. Then the adhered conidia were desorbed, concentrated, and separated using a combination of tITP and MEKC. Finally, the fractions containing the separated conidia were collected from the capillary and analyzed by MALDI-TOF MS. The adhesion efficiency under the optimized experimental conditions was about 80%. This rapid diagnosis will contribute to timely initiation of therapy and increase the patient's chances of survival.