{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,29]],"date-time":"2026-05-29T18:25:37Z","timestamp":1780079137987,"version":"3.54.0"},"reference-count":60,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2024,2,1]],"date-time":"2024-02-01T00:00:00Z","timestamp":1706745600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Health Institute Carlos III","award":["PI19CIII\/00029"],"award-info":[{"award-number":["PI19CIII\/00029"]}]},{"name":"Health Institute Carlos III","award":["FI20CIII\/00002"],"award-info":[{"award-number":["FI20CIII\/00002"]}]},{"name":"Health Institute Carlos III","award":["PTQ018-009754"],"award-info":[{"award-number":["PTQ018-009754"]}]},{"name":"Health Institute Carlos III","award":["CD19CIII\/00011"],"award-info":[{"award-number":["CD19CIII\/00011"]}]},{"name":"Spanish Ministry of Science, Innovation and Universities","award":["PI19CIII\/00029"],"award-info":[{"award-number":["PI19CIII\/00029"]}]},{"name":"Spanish Ministry of Science, Innovation and Universities","award":["FI20CIII\/00002"],"award-info":[{"award-number":["FI20CIII\/00002"]}]},{"name":"Spanish Ministry of Science, Innovation and Universities","award":["PTQ018-009754"],"award-info":[{"award-number":["PTQ018-009754"]}]},{"name":"Spanish Ministry of Science, Innovation and Universities","award":["CD19CIII\/00011"],"award-info":[{"award-number":["CD19CIII\/00011"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>Cryptosporidium spp., Giardia duodenalis and Entamoeba histolytica are species of protozoa- causing diarrhoea that are common worldwide, while Entamoeba dispar, Dientamoeba fragilis and Blastocystis sp. appear to be commensal parasites whose role in pathogenicity remains controversial. We conducted the clinical evaluation of five singleplex and one duplex CerTest VIASURE Real-Time PCR Assays against a large panel of positive DNA samples (n = 358), and specifically to Cryptosporidium spp. (n = 96), G. duodenalis (n = 115), E. histolytica (n = 25) E. dispar (n = 11), Blastocystis sp. (n = 42), D. fragilis (n = 37), and related parasitic phylum species such as Apicomplexa, Euglenozoa, Microsporidia and Nematoda. DNA samples were obtained from clinical stool specimens or cultured isolates in a national reference centre. Estimated diagnostic sensitivity and specificity values were 0.94\u20131 for Cryptosporidium spp., 0.96\u20130.99 for G. duodenalis, 0.96\u20131 for E. histolytica, 1\u20131 for E. dispar, and 1\u20130.99 for D. fragilis in the evaluated singleplex assays. In the duplex assay for the simultaneous detection of Blastocystis sp. and D. fragilis these values were 1\u20130.98 and 1\u20130.99, respectively. Measures of diagnostic precision for repeatability and reproducibility were found to be under acceptable ranges. The assays identified six Cryptosporidium species (C. hominis, C. parvum, C. canis, C. felis, C. scrofarum, and C. ryanae), four G. duodenalis assemblages (A, B, C, and F), and six Blastocystis subtypes (ST1-ST5, and ST8). The evaluated singleplex and duplex VIASURE Real-Time PCR assays provide sensitive, practical, and cost-effective choices to the molecular diagnosis of the main diarrhoea-causing intestinal protists in clinical microbiology and research laboratories.<\/jats:p>","DOI":"10.3390\/diagnostics14030319","type":"journal-article","created":{"date-parts":[[2024,2,1]],"date-time":"2024-02-01T09:04:21Z","timestamp":1706778261000},"page":"319","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Evaluation of the Use of Singleplex and Duplex CerTest VIASURE Real-Time PCR Assays to Detect Common Intestinal Protist Parasites"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8707-5731","authenticated-orcid":false,"given":"Alejandro","family":"Dashti","sequence":"first","affiliation":[{"name":"Parasitology Reference and Research Laboratory, National Centre for Microbiology, Health Institute Carlos III, 28220 Majadahonda, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2742-8827","authenticated-orcid":false,"given":"Henar","family":"Alonso","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Paediatrics, Radiology, and Public Health, Faculty of Medicine, University of Zaragoza, 50009 Saragossa, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6132-893X","authenticated-orcid":false,"given":"Cristina","family":"Escolar-Mi\u00f1ana","sequence":"additional","affiliation":[{"name":"Department of Animal Production and Food Science, Faculty of Veterinary, University of Zaragoza, 50013 Saragossa, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5963-8824","authenticated-orcid":false,"given":"Pamela C.","family":"K\u00f6ster","sequence":"additional","affiliation":[{"name":"Parasitology Reference and Research Laboratory, National Centre for Microbiology, Health Institute Carlos III, 28220 Majadahonda, Spain"},{"name":"Faculty of Health Sciences, Alfonso X El Sabio University (UAX), 28691 Villanueva de la Ca\u00f1ada, Spain"},{"name":"Faculty of Medicine, Alfonso X El Sabio University (UAX), 28691 Villanueva de la Ca\u00f1ada, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bego\u00f1a","family":"Bailo","sequence":"additional","affiliation":[{"name":"Parasitology Reference and Research Laboratory, National Centre for Microbiology, Health Institute Carlos III, 28220 Majadahonda, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4015-8553","authenticated-orcid":false,"given":"David","family":"Carmena","sequence":"additional","affiliation":[{"name":"Parasitology Reference and Research Laboratory, National Centre for Microbiology, Health Institute Carlos III, 28220 Majadahonda, Spain"},{"name":"CIBER Infectious Diseases (CIBERINFEC), Health Institute Carlos III, 28008 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5083-8854","authenticated-orcid":false,"given":"David","family":"Gonz\u00e1lez-Barrio","sequence":"additional","affiliation":[{"name":"Parasitology Reference and Research Laboratory, National Centre for Microbiology, Health Institute Carlos III, 28220 Majadahonda, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,2,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"634","DOI":"10.1128\/CMR.00017-09","article-title":"Clinical significance of enteric protozoa in the immunosuppressed human population","volume":"22","author":"Stark","year":"2009","journal-title":"Clin. 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