{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T16:02:21Z","timestamp":1791388941461,"version":"4.3.3"},"reference-count":20,"publisher":"MDPI AG","issue":"22","license":[{"start":{"date-parts":[[2025,11,19]],"date-time":"2025-11-19T00:00:00Z","timestamp":1763510400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003196","name":"Italian Ministry of Health","doi-asserted-by":"publisher","award":["Ricerca Corrente L3P2"],"award-info":[{"award-number":["Ricerca Corrente L3P2"]}],"id":[{"id":"10.13039\/501100003196","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>Background\/Objectives: The timely diagnosis of schistosomiasis and strongyloidiasis is important because of their potentially severe, even lethal, consequences. European diagnostic laboratories must comply with the European In Vitro Diagnostic (IVD) Regulation, which requires justifying the use of in-house assays when CE-IVD-marked kits are available. We aimed to compare the performance of the Biosynex Helminths AMPLIQUICK\u00ae RT-PCR and the multiplex in-house RT-PCR for the diagnosis of Schistosoma mansoni and Strongyloides stercoralis currently used in our department, an Italian reference centre for tropical diseases. Methods: We conducted a performance comparison study on biobanked frozen stool samples classified as cases or controls according to PCR and\/or copromicroscopy at diagnosis. Both RT-PCRs were performed on DNA re-extracted from the same stool aliquot. Sensitivity and specificity were compared using McNemar\u2019s Chi-squared test, while agreement was assessed using Gwet\u2019s AC1 and Cohen\u2019s K coefficients, and Bland\u2013Altman analysis. Results: A total of 45 S. mansoni cases with 52 controls and 29 S. stercoralis cases with 54 controls were analyzed. For both S. mansoni and S. stercoralis, sensitivity and specificity were not significantly different between RT-PCRs (p = 1). Concordance was perfect for controls (AC1 = 1) in both cohorts, but was poor for S. mansoni cases (AC1 = 0.38) and good for S. stercoralis cases (AC1 = 0.78). Conclusions: Performance was not significantly different between in-house and Biosynex RT-PCRs. Nevertheless, careful assessment of the specific molecular targets included in the panels and prospective evaluation of any newly introduced tests should be implemented to minimize the impact of clinically significant discrepancies.<\/jats:p>","DOI":"10.3390\/diagnostics15222928","type":"journal-article","created":{"date-parts":[[2025,11,20]],"date-time":"2025-11-20T09:44:53Z","timestamp":1763631893000},"page":"2928","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Performance Comparison of In-House and Commercial Biosynex Helmints AMPLIQUICK\u00ae Real-Time PCR Assays for the Diagnosis of Schistosoma mansoni and Strongyloides stercoralis in Stool Samples"],"prefix":"10.3390","volume":"15","author":[{"given":"Davide","family":"Treggiari","sequence":"first","affiliation":[{"name":"Department of Infectious-Tropical Diseases and Microbiology, IRCCS Sacro Cuore Don Calabria Hospital, 37024 Negrar di Valpolicella, VR, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0478-3914","authenticated-orcid":false,"given":"Francesca","family":"Tamarozzi","sequence":"additional","affiliation":[{"name":"Department of Infectious-Tropical Diseases and Microbiology, IRCCS Sacro Cuore Don Calabria Hospital, 37024 Negrar di Valpolicella, VR, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3203-0872","authenticated-orcid":false,"given":"Fabio","family":"Formenti","sequence":"additional","affiliation":[{"name":"Department of Infectious-Tropical Diseases and Microbiology, IRCCS Sacro Cuore Don Calabria Hospital, 37024 Negrar di Valpolicella, VR, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2956-8997","authenticated-orcid":false,"given":"Salvatore","family":"Scarso","sequence":"additional","affiliation":[{"name":"Department of Infectious-Tropical Diseases and Microbiology, IRCCS Sacro Cuore Don Calabria Hospital, 37024 Negrar di Valpolicella, VR, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barbara","family":"Pajola","sequence":"additional","affiliation":[{"name":"Department of Infectious-Tropical Diseases and Microbiology, IRCCS Sacro Cuore Don Calabria Hospital, 37024 Negrar di Valpolicella, VR, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lavinia","family":"Nicolini","sequence":"additional","affiliation":[{"name":"Department of Infectious-Tropical Diseases and Microbiology, IRCCS Sacro Cuore Don Calabria Hospital, 37024 Negrar di Valpolicella, VR, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4561-3447","authenticated-orcid":false,"given":"Cristina","family":"Mazzi","sequence":"additional","affiliation":[{"name":"Clinical Research Unit, IRCCS Sacro Cuore Don Calabria Hospital, 37024 Negrar di Valpolicella, VR, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3933-3892","authenticated-orcid":false,"given":"Francesca","family":"Perandin","sequence":"additional","affiliation":[{"name":"Department of Infectious-Tropical Diseases and Microbiology, IRCCS Sacro Cuore Don Calabria Hospital, 37024 Negrar di Valpolicella, VR, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,11,19]]},"reference":[{"key":"ref_1","unstructured":"World Health Organization (2020). Ending the Neglect to Attain the Sustainable Development Goals: A Road map for Neglected Tropical Diseases 2021\u20132030, World Health Organization."},{"key":"ref_2","unstructured":"(2025, October 02). Status of Schistosomiasis Endemic Countries: 2023. Available online: https:\/\/apps.who.int\/neglected_diseases\/ntddata\/sch\/sch.html."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"658","DOI":"10.1016\/S0140-6736(24)02814-9","article-title":"Human schistosomiasis","volume":"405","author":"Buonfrate","year":"2025","journal-title":"Lancet"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Buonfrate, D., Bisanzio, D., Giorli, G., Odermatt, P., F\u00fcrst, T., Greenaway, C., French, M., Reithinger, R., Gobbi, F., and Montresor, A. (2020). The global prevalence of Strongyloides stercoralis infection. Pathogens, 9.","DOI":"10.3390\/pathogens9060468"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Buonfrate, D., Bradbury, R.S., Watts, M.R., and Bisoffi, Z. (2023). Human strongyloidiasis: Complexities and pathways forward. Clin. Microbiol. Rev., 36.","DOI":"10.1128\/cmr.00033-23"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"102823","DOI":"10.1016\/j.tmaid.2025.102823","article-title":"Prevalence of neglected tropical diseases among migrants living in Europe: A systematic review and meta-analysis","volume":"64","author":"Guido","year":"2025","journal-title":"Travel. Med. Infect. Dis."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"taz075","DOI":"10.1093\/jtm\/taz075","article-title":"Delay in schistosomiasis diagnosis and treatment: A multicenter cohort study in Italy","volume":"27","author":"Comelli","year":"2020","journal-title":"J. Travel. Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/bs.apar.2022.07.001","article-title":"The diagnosis of human and companion animal Strongyloides stercoralis infection: Challenges and solutions. A scoping review","volume":"118","author":"Buonfrate","year":"2022","journal-title":"Adv. Parasitol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"529","DOI":"10.1016\/j.cmi.2015.03.014","article-title":"New diagnostic tools in schistosomiasis","volume":"21","author":"Utzinger","year":"2015","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_10","unstructured":"(2025, October 02). Regulation (EU) 2017\/746 of the European Parliament and of the Council of 5 April 2017 on in Vitro Diagnostic Medical Devices and Repealing Directive 98\/79\/EC and Commission Decision 2010\/227\/EU (Text with EEA Relevance). Available online: https:\/\/eur-lex.europa.eu\/eli\/reg\/2017\/746\/oj\/eng."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"h5527","DOI":"10.1136\/bmj.h5527","article-title":"STARD 2015: An Updated List of Essential Items for Reporting Diagnostic Accuracy Studies","volume":"351","author":"Bossuyt","year":"2015","journal-title":"BMJ"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"625","DOI":"10.1179\/136485908X337490","article-title":"Application of a circulating-cathodic-antigen (CCA) strip test and real-time PCR, in comparison with microscopy, for the detection of Schistosoma haematobium in urine samples from Ghana","volume":"102","author":"Obeng","year":"2008","journal-title":"Ann. Trop. Med. Parasitol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"342","DOI":"10.1016\/j.trstmh.2008.12.001","article-title":"Molecular diagnosis of Strongyloides stercoralis in faecal samples using real-time PCR","volume":"103","author":"Verweij","year":"2009","journal-title":"Trans. R. Soc. Trop. Med. Hyg."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1186\/s41182-024-00619-y","article-title":"Prevalence of Strongyloides stercoralis and other helminths in four districts of Madagascar","volume":"52","author":"Scarso","year":"2024","journal-title":"Trop. Med. Health"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"e169","DOI":"10.1093\/infdis\/jiaf237","article-title":"Improving molecular epidemiological surveillance of strongyloidiasis upon differentiation of Strongyloides fuelleborni fuelleborni from Strongyloides stercoralis","volume":"232","author":"Cunningham","year":"2025","journal-title":"J. Infect. Dis."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Akenten, C.W., Weinreich, F., Paintsil, E.K., Amuasi, J., Fosu, D., Loderst\u00e4dt, U., May, J., Frickmann, H., and Dekker, D. (2022). Intestinal helminth infections in Ghanaian children from the Ashanti region between 2007 and 2008\u2014A retrospective cross-sectional real-time PCR-Based assessment. Trop. Med. Infect. Dis., 7.","DOI":"10.3390\/tropicalmed7110374"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"S3","DOI":"10.1016\/S1386-6532(02)00197-X","article-title":"Clinical virology in real time","volume":"25","author":"Niesters","year":"2002","journal-title":"J. Clin. Virol."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Llewellyn, S., Inpankaew, T., Nery, S.V., Gray, D.J., Verweij, J.J., Clements, A.C., Gomes, S.J., Traub, R., and McCarthy, J.S. (2016). Application of a multiplex quantitative PCR to assess prevalence and intensity of intestinal parasite infections in a controlled clinical trial. PLoS Negl. Trop. Dis., 10.","DOI":"10.1371\/journal.pntd.0004380"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1348\/000711006X126600","article-title":"Computing inter-rater reliability and its variance in the presence of high agreement","volume":"61","author":"Gwet","year":"2008","journal-title":"British J. Math. Stat. Psychol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1240","DOI":"10.4269\/ajtmh.21-0803","article-title":"Sensitive diagnosis and post-treatment follow-up of Schistosoma mansoni infections in asymptomatic Eritrean refugees by circulating anodic antigen detection and polymerase chain reaction","volume":"106","author":"Hoekstra","year":"2022","journal-title":"Am. J. Trop. Med. Hyg."}],"container-title":["Diagnostics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-4418\/15\/22\/2928\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,24]],"date-time":"2025-11-24T10:40:46Z","timestamp":1763980846000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-4418\/15\/22\/2928"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,19]]},"references-count":20,"journal-issue":{"issue":"22","published-online":{"date-parts":[[2025,11]]}},"alternative-id":["diagnostics15222928"],"URL":"https:\/\/doi.org\/10.3390\/diagnostics15222928","relation":{},"ISSN":["2075-4418"],"issn-type":[{"value":"2075-4418","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,11,19]]}}}