{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T21:57:23Z","timestamp":1791237443113,"version":"4.1.0"},"reference-count":40,"publisher":"Wildlife Disease Association","issue":"1","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Journal of Wildlife Diseases"],"published-print":{"date-parts":[[2026,2,16]]},"abstract":"<jats:p>The collection and storage of swab samples for molecular diagnostics is a routine component of wildlife health surveillance. The suitability of different sample storage conditions for maximizing the recovery of pathogen DNA in most species has not been assessed; therefore, the aim of this study was to identify a preferred storage method for swabs collected for the detection of frog virus 3 (FV3), a significant chelonian pathogen. Sterile swabs were inoculated in triplicate with a plasmid containing known quantities of FV3 DNA from 10\u00b0 to 107 copies. Swabs were then stored under one of the following four conditions: 1) dry frozen at \u201320 \u00b0C; 2) immersed in phosphate-buffered saline (PBS) and frozen at \u201320 \u00b0C; 3) immersed in a nucleic acid preservative (RNAlater) and frozen at \u201320 \u00b0C; and 4) immersed in 100% ethanol at ambient room temperature. Swabs remained undisturbed under designated storage conditions for 14 d, at which point DNA extraction and conventional and quantitative PCR for FV3 detection were performed. Conventional PCR amplified down to the lowest expected FV3 target copy number (10,000 copies\/swab) for the dry-frozen and PBS-frozen treatment groups. Conventional PCR amplification was inconsistent for the ethanol and RNAlater treatment groups. Quantitative PCR on dry-frozen samples successfully amplified as low as 100 FV3 target copies\/swab with a mean recovery of 90%, with all other storage methods amplifying only down to 10,000 copies\/swab. Findings suggest there is improved detection of pathogen DNA for samples stored from collection to extraction under the dry-frozen method. Swab sample storage recommendations for future applications should be observed within the context of study-specific objectives and target pathogens. Furthermore, failure to detect fewer than 100 copies\/swab of FV3 from any storage method may have clinically significant ramifications and suggests that the differences in DNA recovery based on extraction method should also be examined.<\/jats:p>","DOI":"10.7589\/jwd-d-24-00165","type":"journal-article","created":{"date-parts":[[2026,2,16]],"date-time":"2026-02-16T09:34:48Z","timestamp":1771234488000},"page":"154-160","source":"Crossref","is-referenced-by-count":1,"title":["INVESTIGATION OF SAMPLE STORAGE CONDITIONS FOR OPTIMIZED DETECTION OF FROG VIRUS 3 DNA IN CHELONIAN SWAB SAMPLES"],"prefix":"10.7589","volume":"62","author":[{"given":"Carly R.","family":"Harkey","sequence":"first","affiliation":[{"name":"Wildlife Epidemiology Laboratory, College of Veterinary Medicine, University of Illinois, 2001 S. Lincoln Ave., Urbana, Illinois 61802, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Laura A.","family":"Adamovicz","sequence":"additional","affiliation":[{"name":"Wildlife Epidemiology Laboratory, College of Veterinary Medicine, University of Illinois, 2001 S. Lincoln Ave., Urbana, Illinois 61802, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maris J.","family":"Daleo","sequence":"additional","affiliation":[{"name":"Wildlife Epidemiology Laboratory, College of Veterinary Medicine, University of Illinois, 2001 S. Lincoln Ave., Urbana, Illinois 61802, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amber L.","family":"Simmons","sequence":"additional","affiliation":[{"name":"Wildlife Epidemiology Laboratory, College of Veterinary Medicine, University of Illinois, 2001 S. Lincoln Ave., Urbana, Illinois 61802, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew C.","family":"Allender","sequence":"additional","affiliation":[{"name":"Wildlife Epidemiology Laboratory, College of Veterinary Medicine, University of Illinois, 2001 S. Lincoln Ave., Urbana, Illinois 61802, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"4415","reference":[{"key":"15","doi-asserted-by":"crossref","unstructured":"VL Hale, CL Tan, R Knight, et al.  \"J Microbiol Methods.\"  113, \"Effect of preservation method on spider monkey (Ateles geoffroyi) fecal microbiota over 8 weeks.\" 16 (2015)","DOI":"10.1016\/j.mimet.2015.03.021"},{"key":"33","doi-asserted-by":"crossref","unstructured":"AK Schilling, MV Mazzamuto, C Romeo,  \"Animals (Basel).\"  12, \"A review of non-invasive sampling in wildlife disease and health research: What's new?\" 1719 (2022)","DOI":"10.3390\/ani12131719"},{"key":"38","doi-asserted-by":"crossref","unstructured":"TJ Willis, M Kelly, L Adamovicz, et al.  \"J Wildl Dis.\"  58, \"Enhancing chelonian health assessments by determining agreement in pathogen detection between sampling sites using quantitative PCR.\" 399 (2022)","DOI":"10.7589\/JWD-D-21-00096"},{"key":"3","doi-asserted-by":"crossref","unstructured":"BH Bach, AB Quigley, KM Gaynor, et al.  \"Mamm Biol.\"  102, \"Identifying individual ungulates from fecal DNA: A comparison of field collection methods to maximize efficiency, ease, and success.\" 863 (2022)","DOI":"10.1007\/s42991-021-00176-5"},{"key":"4","unstructured":"JL Belant  and AR Deese  \"Hum Wildl Interact.\"  4, \"Importance of wildlife disease surveillance.\" 165 (2010)"},{"key":"5","doi-asserted-by":"crossref","unstructured":"LM Bergner, RJ Orton, A da Silva Filipe, et al.  \"Mol Ecol Resour.\"  19, \"Using noninvasive metagenomics to characterize viral communities from wildlife.\" 128 (2019)","DOI":"10.1111\/1755-0998.12946"},{"key":"6","doi-asserted-by":"crossref","unstructured":"KH Bodawatta, K Puzejova, K Sam, et al.  \"Anim Microbiome.\"  2, \"Cloacal swabs and alcohol bird specimens are good proxies for compositional analyses of gut microbial communities of Great tits (Parus major).\" 9 (2020)","DOI":"10.1186\/s42523-020-00026-8"},{"key":"8","doi-asserted-by":"crossref","unstructured":"JJ Campanella  and JV Smalley  \"BMC Genet.\"  7, \"A minimally invasive method of piscine tissue collection and an analysis of long-term field-storage conditions for samples.\" 32 (2006)","DOI":"10.1186\/1471-2156-7-32"},{"key":"9","doi-asserted-by":"crossref","unstructured":"JM Choo, LEX Leong, GB Rogers,  \"Sci Rep.\"  5, \"Sample storage conditions significantly influence faecal microbiome profiles.\" 16350 (2015)","DOI":"10.1038\/srep16350"},{"key":"11","doi-asserted-by":"crossref","unstructured":"SH Feldman, J Wimsatt, RE Marchang, et al.  \"J Wild Dis.\"  2, \"A novel mycoplasma detected in association with upper respiratory disease syndrome in free-ranging eastern box turtles (Terrapene carolina carolina) in Virginia.\" 279 (2006)","DOI":"10.7589\/0090-3558-42.2.279"},{"key":"22","doi-asserted-by":"crossref","unstructured":"PD Kirkland  and MJ Frost  \"Pathology.\"  52, \"The impact of viral transport media on PCR assay results for the detection of nucleic acid from SARS-CoV-2.\" 811 (2020)","DOI":"10.1016\/j.pathol.2020.09.013"},{"key":"23","doi-asserted-by":"crossref","unstructured":"D. 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free-ranging Blanding's turtles (Emydoidea blandingii) from Illinois.\" 547 (2019)","DOI":"10.1638\/2018-0074"},{"key":"36","doi-asserted-by":"crossref","unstructured":"RR Sim, RJ Ossiboff, J Nelson, et al.  \"J Zoo Wildl Med.\"  52, \"Emydid herpesvirus 1 infections in western pond turtles (Actinemys marmorata) and a red-eared slider (Trachemys scripta elegans) with fatal and nonfatal outcomes.\" 1275 (2021)","DOI":"10.1638\/2021-0044"},{"key":"31","unstructured":"\"R: A Language and Environment for Statistical Computing.\"  (2023)"}],"container-title":["Journal of Wildlife Diseases"],"original-title":[],"deposited":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T21:18:50Z","timestamp":1791235130000},"score":1,"resource":{"primary":{"URL":"https:\/\/bioone.org\/journals\/journal-of-wildlife-diseases\/volume-62\/issue-1\/JWD-D-24-00165\/INVESTIGATION-OF-SAMPLE-STORAGE-CONDITIONS-FOR-OPTIMIZED-DETECTION-OF-FROG\/10.7589\/JWD-D-24-00165.full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2,16]]},"references-count":40,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026,1,1]]}},"URL":"https:\/\/doi.org\/10.7589\/jwd-d-24-00165","relation":{},"ISSN":["0090-3558","1943-3700"],"issn-type":[{"value":"0090-3558","type":"print"},{"value":"1943-3700","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,16]]}}}