Next Generation Sequencing for miRNA Detection on the Exhaled Breath Condensate: A Pilot Study.

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Centre for Occupational and Environmental Health, Division of Population Health, Health Services Research & Primary Care, University of Manchester, Manchester, UK.

Published: April 2023

Introduction: Exhaled breath condensate (EBC) sampling has been suggested as a less-invasive and cost-effective method to detect biological macromolecules, including miRNA. To explore the feasibility of its use as a biomarker of early effects of asbestos exposure, we conducted a preliminary test on male volunteers by comparing the miRNA profile in the EBC and the plasma using 2 different sequencing platforms.

Methods: Six male volunteers, all retired and unexposed to dust or fumes, participated in the test. RNA was extracted from 200 μL EBC samples and same-size plasma samples. Sample aliquots were processed in 2 laboratories using 2 different sequencing platforms: a MiSeq Illumina platform and a more performing HiSeq Illumina platform.

Results: The HiSeq3000 sequencing platform identified twice as many unique molecular indexes (UMI)-validated miRNA as the MiSeq platform. The Spearman's correlation coefficient between EBC counts and plasma counts was significant in 5/6 subjects with either platform (MiSeq = 0.128-0.508,  = .026-<.001; HiSeq = 0.156-0.412,  = .001-<.001). The intraclass correlation coefficient confirmed the consistency of the miRNA profile over the 6 participants with both biospecimens. Exploring the agreement between the EBC and plasma samples with Bland-Altman plots showed that using the HiSeq3000 platform substantially improved the EBC miRNA detection rate.

Conclusion: Our preliminary study confirms that, when using the HiSeq sequencing platform, EBC sampling is a suitable, non-invasive method to detect the miRNA profile in healthy subjects.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10070752PMC
http://dx.doi.org/10.1177/25168657231160985DOI Listing

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