Detection of pulmonary amylase activity in exhaled breath condensate

Zweifel, Martin; Rechsteiner, T.; Hofer, M.; Boehler, A. (2013). Detection of pulmonary amylase activity in exhaled breath condensate. Journal of breath research, 7(4), 046007. IOP Science 10.1088/1752-7155/7/4/046007

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Amylase activity in exhaled breath condensate (EBC) is usually interpreted as an indication of oropharyngeal contamination despite the fact that amylase can be found in pulmonary excretions. The aim of this study was to recruit and refine an amylase assay in order to detect amylase activity in any EBC sample and to develop a method to identify EBC samples containing amylase of pulmonary origin. EBC was collected from 40 volunteers with an EcoScreen condenser. Amylase assays and methods to discriminate between oropharyngeal and pulmonary proteins were tested and developed using matched EBC and saliva samples. Our refined 2-chloro-4-nitrophenyl-α-D-maltotriosid (CNP-G3) assay was 40-fold more sensitive than the most sensitive commercial assay and allowed detection of amylase activity in 30 µl of EBC. We developed a dot-blot assay which allowed detection of salivary protein in saliva diluted up to 150 000-fold. By plotting amylase activity against staining intensity we identified a few EBC samples with high amylase activity which were aligned with diluted saliva. We believe that EBC samples aligned with diluted saliva contain amylase activity introduced during EBC collection and that all other EBC samples contain amylase activity of pulmonary origin and are basically free of oropharyngeal protein contamination.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Department of Haematology, Oncology, Infectious Diseases, Laboratory Medicine and Hospital Pharmacy (DOLS) > Clinic of Medical Oncology

UniBE Contributor:

Zweifel, Martin

Subjects:

600 Technology > 610 Medicine & health

ISSN:

1752-7163

Publisher:

IOP Science

Language:

English

Submitter:

Marianne Zahn

Date Deposited:

12 May 2014 11:40

Last Modified:

05 Dec 2022 14:30

Publisher DOI:

10.1088/1752-7155/7/4/046007

PubMed ID:

24287549

URI:

https://boris.unibe.ch/id/eprint/45051

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