ArticleJournal of cystic fibrosis : official journal of the European Cystic Fibrosis Society2015
Implications of multiple freeze-thawing on respiratory samples for culture-independent analyses.
Article in Journal of cystic fibrosis : official journal of the European Cystic Fibrosis Society, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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Who cites it
27 citing papers in PubMed.
- Remodelling of cystic fibrosis respiratory microbiota in response to extended elexacaftor-tezacaftor-ivacaftor therapy.Microbiome · 2026Article
- Analytic attributes of the 16S rRNA gene sequencing methodology for human gut microbiota characterization.Applied microbiology and biotechnology · 2026Article
- Effects of heat-assisted sample desiccation on microbiome surveys.Environmental microbiome · 2026Article
- Effects of repeated freeze and thaw cycles on the stability of faecal microbiome composition.Scientific reports · 2026Article
- Advancing diagnostics in suspected periprosthetic joint infections using synthetic synovial fluid and microcalorimetry.Journal of bone and joint infection · 2026Article
- Effects of Vacuum-Heat-Assisted Sample Desiccation on Microbiome Surveys.Molecular ecology resources · 2025Article
- Species turnover within cystic fibrosis lung microbiota is indicative of acute pulmonary exacerbation onset.Microbiome · 2025Article
- Effects of Different Preservation Methods on the Structure and Diversity of Intestinal Microbiota of Marine Fishes.Current microbiology · 2025Article
- Subtracting the background by reducing cell-free DNA's confounding effects on Mycobacterium tuberculosis quantitation and the sputum microbiome.Scientific reports · 2024Article
- The Metabolic Potential of the Human Lung Microbiome.Microorganisms · 2024Article
- Ecological patterns and processes of temporal turnover within lung infection microbiota.Microbiome · 2024Article
- Sputum handling for rheology.Scientific reports · 2023Article
- Bacterial Culture Underestimates Lung Pathogen Detection and Infection Status in Cystic Fibrosis.Microbiology spectrum · 2022Article
- Towards standardized and reproducible research in skin microbiomes.Environmental microbiology · 2022Review
- The microbiome of common bedding materials before and after use on commercial dairy farms.Animal microbiome · 2022Article
- Lung function and microbiota diversity in cystic fibrosis.Microbiome · 2020Article
- The Mycobiome in Health and Disease: Emerging Concepts, Methodologies and Challenges.Mycopathologia · 2020Review
- Systematic Analysis of Impact of Sampling Regions and Storage Methods on Fecal Gut Microbiome and Metabolome Profiles.mSphere · 2020Article
- Understanding and overcoming the pitfalls and biases of next-generation sequencing (NGS) methods for use in the routine clinical microbiological diagnostic laboratory.European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology · 2019Review
- 'Candidatus Ornithobacterium hominis': insights gained from draft genomes obtained from nasopharyngeal swabs.Microbial genomics · 2019Article
Corrections and comments
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Authors and funding
9 authors.
Funding
Abstract
backgroundBest practice when performing culture-independent microbiological analysis of sputum samples involves their rapid freezing and storage at -80°C. However, accessing biobanked collections can mean that material has been passed through repeated freeze-thaw cycles. The aim of this study was to determine the impact of these cycles on microbial community profiles.
methodsSputum was collected from eight adults with cystic fibrosis, and each sample was subjected to six freeze-thaw cycles. Following each cycle, an aliquot was removed and treated with propidium monoazide (PMA) prior to DNA extraction and 16S rRNA gene pyrosequencing.
resultsThe impact of freeze-thaw cycles was greatest on rare members of the microbiota, with variation beyond that detected with within-sample repeat analysis observed after three cycles.
conclusionFour or more freeze thaw cycles result in a significant distortion of microbiota profiles from CF sputum.
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