{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T02:11:28Z","timestamp":1790475088421,"version":"4.1.0"},"reference-count":32,"publisher":"Public Library of Science (PLoS)","issue":"2","license":[{"start":{"date-parts":[[2021,2,17]],"date-time":"2021-02-17T00:00:00Z","timestamp":1613520000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"French Ministry of Health"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:sec id=\"sec001\"><jats:title>Introduction<\/jats:title><jats:p><jats:italic>Toxoplasma<\/jats:italic>-PCR is essential to diagnose ocular, cerebral, disseminated and congenital toxoplasmosis. This multicenter study evaluated the impact of sample storage duration at +4\u00b0C on PCR assay performances in order to propose guidelines for the storage of samples during shipment or\/and before PCR.<\/jats:p><\/jats:sec><jats:sec id=\"sec002\"><jats:title>Materials and methods<\/jats:title><jats:p>Five matrices, amniotic (AF), cerebrospinal (CSF), and bronchoalveolar lavage fluids (BALF), whole blood (WB) and buffy coat (BC), were artificially spiked with different amounts of<jats:italic>Toxoplasma gondii<\/jats:italic>(20, 100, 500 tachyzoites per mL of sample) or with previously infected THP1 cells. DNA extractions were performed at day 0 and after 2, 4 and 7 days of storage at +4\u00b0C. Each extract was amplified at least twice by real-time PCR.<\/jats:p><\/jats:sec><jats:sec id=\"sec003\"><jats:title>Results<\/jats:title><jats:p>A total of 252 spiked samples was studied. No increase of crossing point was observed and all samples were positive for AF, BALF, BC and infected THP1-spiked WB after up to 7 days at 4\u00b0C. For CSF spiked with 20 parasites\/mL, only 50% of PCR reactions were positive at D7 (p&lt;0.05). For WB spiked with type II parasites, all reactions remained positive at D7 but amplifications were significantly delayed from D2; and for WB spiked with RH strain, the proportion of positive reactions decreased at D7.<\/jats:p><\/jats:sec><jats:sec id=\"sec004\"><jats:title>Conclusion<\/jats:title><jats:p>The storage of clinical samples at +4\u00b0C is compatible with the molecular detection of<jats:italic>T<\/jats:italic>.<jats:italic>gondii<\/jats:italic>parasites. Provided that PCR assays are performed in duplicate, storage of samples is possible up to 7 days. However, from the fifth day onwards, and for samples susceptible to contain low parasitic loads, we recommend to perform the PCR in multiplicate.<\/jats:p><\/jats:sec>","DOI":"10.1371\/journal.pone.0246802","type":"journal-article","created":{"date-parts":[[2021,2,17]],"date-time":"2021-02-17T18:41:58Z","timestamp":1613587318000},"page":"e0246802","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":6,"title":["Impact of pre-analytic step duration on molecular diagnosis of toxoplasmosis for five types of biological 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