{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,28]],"date-time":"2026-01-28T05:24:21Z","timestamp":1769577861143,"version":"3.49.0"},"reference-count":38,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2021,11,10]],"date-time":"2021-11-10T00:00:00Z","timestamp":1636502400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Certest Biotec SL.","award":["not applicable"],"award-info":[{"award-number":["not applicable"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Nowadays, many commercial kits allowing the detection of digestive parasites by DNA amplification methods have been developed, including simplex PCR assays (SimpPCRa) allowing the identification of a single parasite, and multiplex PCR assays (MultPCRa) allowing the identification of several parasites at once. Thus, aimed at improving the diagnosis of intestinal protozoal infections, it is essential to evaluate the performances of these new tools. A total of 174 DNA samples collected between 2007 and 2017 were retrospectively included in this study. Performances of four commercial SimpPCRa (i.e., CerTest-VIASURETM) and three MultPCRa (i.e., CerTest-VIASURETM, FAST-TRACK-Diagnostics-FTD-Stool-ParasiteTM and DIAGENODE-Gastroenteritis\/Parasite-panel-ITM) were evaluated for the detection of Cryptosporidium spp., Entamoeba spp., and Giardia intestinalis in stool samples compared to our routinely used in-house SimpPCRa. Globally, the SimpPCRa showed better sensitivity\/specificity for the detection of G. intestinalis, E. histolytica, E. dispar, and Cryptosporidium spp. (i.e., 96.9\/93.6%; 100\/100%; 95.5\/100%; and 100\/99.3%, respectively), compared to the three commercial MultPCRa tested. All in all, we showed that MultPCRa offer an interesting alternative for the detection of protozoans in stool samples depending on the clinical context.<\/jats:p>","DOI":"10.3390\/microorganisms9112325","type":"journal-article","created":{"date-parts":[[2021,11,10]],"date-time":"2021-11-10T09:19:21Z","timestamp":1636535961000},"page":"2325","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Commercial Simplex and Multiplex PCR Assays for the Detection of Intestinal Parasites Giardia intestinalis, Entamoeba spp., and Cryptosporidium spp.: Comparative Evaluation of Seven Commercial PCR Kits with Routine In-House Simplex PCR Assays"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4214-8022","authenticated-orcid":false,"given":"Louise","family":"Basmaciyan","sequence":"first","affiliation":[{"name":"Department of Parasitology\/Mycology, University Hospital of Dijon, 21000 Dijon, France"},{"name":"CNR LE Cryptosporidiosis Collaborating Laboratory, Sant\u00e9 Publique France, 21000 Dijon, France"},{"name":"UMR PAM Univ Bourgogne Franche-Comt\u00e9\u2014AgroSup Dijon\u2014Equipe Vin, Aliment, Microbiologie, Stress, 21078 Dijon, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alexandre","family":"Fran\u00e7ois","sequence":"additional","affiliation":[{"name":"Department of Parasitology\/Mycology, University Hospital of Dijon, 21000 Dijon, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anne","family":"Vincent","sequence":"additional","affiliation":[{"name":"Department of Parasitology\/Mycology, University Hospital of Dijon, 21000 Dijon, France"},{"name":"CNR LE Cryptosporidiosis Collaborating Laboratory, Sant\u00e9 Publique France, 21000 Dijon, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"St\u00e9phane","family":"Valot","sequence":"additional","affiliation":[{"name":"Department of Parasitology\/Mycology, University Hospital of Dijon, 21000 Dijon, France"},{"name":"CNR LE Cryptosporidiosis Collaborating Laboratory, Sant\u00e9 Publique France, 21000 Dijon, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alain","family":"Bonnin","sequence":"additional","affiliation":[{"name":"Department of Parasitology\/Mycology, University Hospital of Dijon, 21000 Dijon, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3418-2597","authenticated-orcid":false,"given":"Damien","family":"Costa","sequence":"additional","affiliation":[{"name":"CNR LE Cryptosporidiosis, Sant\u00e9 Publique France, 76000 Rouen, France"},{"name":"Department of Parasitology\/Mycology, University Hospital of Rouen, 76000 Rouen, France"},{"name":"EA ESCAPE 7510, University of Medicine Pharmacy Rouen, 76000 Rouen, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3188-7794","authenticated-orcid":false,"given":"Romy","family":"Razakandrainibe","sequence":"additional","affiliation":[{"name":"CNR LE Cryptosporidiosis, Sant\u00e9 Publique France, 76000 Rouen, France"},{"name":"EA ESCAPE 7510, University of Medicine Pharmacy Rouen, 76000 Rouen, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Florent","family":"Morio","sequence":"additional","affiliation":[{"name":"Laboratorial de Parasitologie-Mycologie, Institut de Biologie, CHU de Nantes, 44000 Nantes, France"},{"name":"D\u00e9partement de Parasitologie et Mycologie M\u00e9dicale, EA1155-IICiMed, Institut de Recherche en Sant\u00e9 2, Universit\u00e9 de Nantes, Nantes Atlantique Universit\u00e9s, 44035 Nantes, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3386-2240","authenticated-orcid":false,"given":"Loic","family":"Favennec","sequence":"additional","affiliation":[{"name":"CNR LE Cryptosporidiosis, Sant\u00e9 Publique France, 76000 Rouen, France"},{"name":"Department of Parasitology\/Mycology, University Hospital of Rouen, 76000 Rouen, France"},{"name":"EA ESCAPE 7510, University of Medicine Pharmacy Rouen, 76000 Rouen, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fr\u00e9d\u00e9ric","family":"Dalle","sequence":"additional","affiliation":[{"name":"Department of Parasitology\/Mycology, University Hospital of Dijon, 21000 Dijon, France"},{"name":"CNR LE Cryptosporidiosis Collaborating Laboratory, Sant\u00e9 Publique France, 21000 Dijon, France"},{"name":"UMR PAM Univ Bourgogne Franche-Comt\u00e9\u2014AgroSup Dijon\u2014Equipe Vin, Aliment, Microbiologie, Stress, 21078 Dijon, France"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,11,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"429","DOI":"10.1007\/s11894-007-0054-8","article-title":"Amebic colitis: New insights into pathogenesis and treatment","volume":"9","author":"Bercu","year":"2007","journal-title":"Curr. Gastroenterol. Rep."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"698","DOI":"10.1016\/j.meegid.2008.05.004","article-title":"Molecular epidemiology of amebiasis","volume":"8","author":"Ali","year":"2008","journal-title":"Infect. Genet."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1023","DOI":"10.1016\/j.meegid.2009.06.008","article-title":"Reassessment of the epidemiology of amebiasis: State of the art","volume":"9","author":"Rojas","year":"2009","journal-title":"Infect. Evol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1128\/CMR.05038-11","article-title":"Enteric protozoa in the developed world: A public health perspective","volume":"25","author":"Fletcher","year":"2012","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"42","DOI":"10.4081\/jphr.2013.e9","article-title":"Prevalence of gastrointestinal pathogens in developed and developing countries: Systematic review and meta-analysis","volume":"2","author":"Fletcher","year":"2013","journal-title":"J. Public Health Res."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1016\/S0140-6736(14)61682-2","article-title":"Global, regional, and national age-sex specific all-cause and cause-specific mortality for 240 causes of death, 1990\u20132013: A systematic analysis for the Global Burden of Disease Study 2013","volume":"385","author":"Abubakar","year":"2015","journal-title":"Lancet"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"20205","DOI":"10.2807\/ese.17.26.20205-en","article-title":"Infectious diseases among travellers and migrants in Europe, EuroTravNet 2010","volume":"17","author":"Gautret","year":"2012","journal-title":"Eurosurveillance"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"328","DOI":"10.1136\/gut.2006.100198","article-title":"Effect of chronic Giardia lamblia infection on epithelial transport and barrier function in human duodenum","volume":"56","author":"Troeger","year":"2007","journal-title":"Gut"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"609","DOI":"10.4269\/ajtmh.2009.80.609","article-title":"Global etiology of travelers\u2019 diarrhea: Systematic review from 1973 to the present","volume":"80","author":"Shah","year":"2009","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_10","first-page":"182","article-title":"L\u2019amoebose intestinale humaine revisit\u00e9e: Entamoeba histolytica, pathog\u00e8ne, est moins fr\u00e9quent que Entamoeba dispar, non pathog\u00e8ne","volume":"22","author":"Thellier","year":"2007","journal-title":"La Lett. L\u2019infectiologue"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S1674-8301(11)60001-8","article-title":"Cryptosporidiosis-an overview","volume":"25","author":"Leitch","year":"2011","journal-title":"J. Biomed. Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"712","DOI":"10.1128\/JCM.02877-13","article-title":"Detection of intestinal protozoa in the clinical laboratory","volume":"52","author":"McHardy","year":"2014","journal-title":"J. Clin. Microbiol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"888","DOI":"10.1136\/jcp.56.12.888","article-title":"Best Practice No 174. Best practice guidelines for the examination of specimens for the diagnosis of parasitic infections in routine diagnostic laboratories","volume":"56","author":"Francis","year":"2003","journal-title":"J. Clin. Pathol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1128\/JCM.41.1.237-241.2003","article-title":"Detection and differentiation of Entamoeba histolytica and Entamoeba dispar isolates in clinical samples by PCR and enzyme-linked immunosorbent assay","volume":"41","author":"Gonin","year":"2003","journal-title":"J. Clin. Microbiol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"680","DOI":"10.1080\/00365540510037812","article-title":"PCR differentiation of Entamoeba histolytica and Entamoeba dispar from patients with amoeba infection initially diagnosed by microscopy","volume":"37","author":"Lebbad","year":"2005","journal-title":"Scand. J. Infect. Dis."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"3196","DOI":"10.1128\/JCM.00778-06","article-title":"Differential detection of Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii by a single-round PCR assay","volume":"44","author":"Hamzah","year":"2006","journal-title":"J. Clin. Microbiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1205","DOI":"10.1016\/j.cmi.2018.02.007","article-title":"Evaluation of a new multiplex PCR assay (ParaGENIE G-Amoeba Real-Time PCR kit) targeting Giardia intestinalis, Entamoeba histolytica and Entamoeba dispar\/Entamoeba moshkovskii from stool specimens: Evidence for the limited performances of microscopy-based approach for amoeba species identification","volume":"24","author":"Morio","year":"2018","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"371","DOI":"10.1128\/CMR.00122-13","article-title":"Molecular testing for clinical diagnosis and epidemiological investigations of intestinal parasitic infections","volume":"27","author":"Verweij","year":"2014","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"3723","DOI":"10.1128\/JCM.02103-15","article-title":"Multiplex molecular panels for the diagnosis of gastrointestinal infection: Performance, result interpretation and cost-effectiveness","volume":"53","author":"Binnicker","year":"2015","journal-title":"J. Clin. Microbiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"173","DOI":"10.4269\/ajtmh.16-0781","article-title":"Performance of real-time polymerase chain reaction assays for the detection of 20 gastrointestinal parasites in clinical samples from Senegal","volume":"97","author":"Sow","year":"2017","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"e1","DOI":"10.1016\/j.cmi.2015.10.019","article-title":"Is real-time PCR-based diagnosis similar in performance to routine parasitological examination for the identification of Giardia intestinalis, Cryptosporidium parvum\/Cryptosporidium hominis and Entamoeba histolytica from stool samples? Evaluation of a new commercial multiplex PCR assay and literature review","volume":"22","author":"Laude","year":"2016","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"188","DOI":"10.2143\/ACB.3170","article-title":"Detection of Giardia lamblia, Cryptosporidium spp. and Entamoeba histolytica in clinical stool samples by using multiplex real-time PCR after automated DNA isolation","volume":"68","author":"Rossen","year":"2013","journal-title":"Acta Clin. Belg."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Paulos, S., Saugar, J.M., de Lucio, A., Fuentes, I., Mateo, M., and Carmena, D. (2019). Comparative performance evaluation of four commercial multiplex real-time PCR assays for the detection of the diarrhoea-causing protozoa Cryptosporidium hominis\/parvum, Giardia duodenalis and Entamoeba histolytica. PLoS ONE, 14.","DOI":"10.1371\/journal.pone.0215068"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1051\/parasite\/2018049","article-title":"Comparison of three commercial multiplex PCR assays for the diagnosis of intestinal protozoa","volume":"25","author":"Autier","year":"2018","journal-title":"Parasite"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"122","DOI":"10.1016\/j.jinf.2013.04.009","article-title":"Systematic application of multiplex PCR enhances the detection of bacteria, parasites, and viruses in stool samples","volume":"67","author":"McAuliffe","year":"2013","journal-title":"J. Infect."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"472","DOI":"10.1128\/JCM.02658-12","article-title":"A laboratory-developed TaqMan Array Card for simultaneous detection of 19 enteropathogens","volume":"51","author":"Liu","year":"2013","journal-title":"J. Clin. Microbiol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1041","DOI":"10.4269\/ajtmh.12-0726","article-title":"A novel, multi-parallel, real-time polymerase chain reaction approach for eight gastrointestinal parasites provides improved diagnostic capabilities to resource-limited at-risk populations","volume":"88","author":"Mejia","year":"2013","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1128\/JCM.01796-10","article-title":"Evaluation of multiplex tandem real-time PCR for detection of Cryptosporidium spp., Dientamoeba fragilis, Entamoeba histolytica, and Giardia intestinalis in clinical stool samples","volume":"49","author":"Stark","year":"2011","journal-title":"J. Clin. Microbiol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1051\/parasite\/2013051","article-title":"Application of the NucliSENS easyMAG system for nucleic acid extraction: Optimization of DNA extraction for molecular diagnosis of parasitic and fungal diseases","volume":"20","author":"Jeddi","year":"2013","journal-title":"Parasite"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1016\/S0890-8508(03)00057-4","article-title":"Real-time PCR for the detection of Giardia lamblia","volume":"17","author":"Verweij","year":"2003","journal-title":"Mol. Cell. Probes"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"305","DOI":"10.1016\/S0035-9203(03)90153-2","article-title":"Overdiagnosis of amoebiasis in the absence of Entamoeba histolytica among patients presenting with diarrhoea in Wonji and Akaki, Ethiopia","volume":"97","author":"Kebede","year":"2003","journal-title":"Trans. R. Soc. Trop. Med. Hyg."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Brunet, J., Lemoine, J.P., Pesson, B., Valot, S., Sautour, M., Dalle, F., Muller, C., Borni-Duval, C., Caillard, S., and Moulin, B. (2016). Ruling out nosocomial transmission of Cryptosporidium in a renal transplantation unit: Case report. BMC Infect. Dis., 16.","DOI":"10.1186\/s12879-016-1661-5"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"980","DOI":"10.4269\/ajtmh.2012.11-0661","article-title":"Evaluation of a rapid point-of-care fecal antigen detection test for Entamoeba histolytica","volume":"86","author":"Korpe","year":"2012","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1220","DOI":"10.1128\/JCM.42.3.1220-1223.2004","article-title":"Simultaneous detection of Entamoeba histolytica, Giardia lamblia, and Cryptosporidium parvum in fecal samples by using multiplex real-time PCR","volume":"42","author":"Verweij","year":"2004","journal-title":"J. Clin. Microbiol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1678","DOI":"10.1128\/JCM.02261-07","article-title":"Comparison of stool antigen detection kits to PCR for diagnosis of amebiasis","volume":"46","author":"Stark","year":"2008","journal-title":"J. Clin. Microbiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2258","DOI":"10.1128\/JCM.00530-06","article-title":"Comparison of a stool antigen detection kit and PCR for diagnosis of Entamoeba histolytica and Entamoeba dispar infections in asymptomatic cyst passers in Iran","volume":"44","author":"Rezaian","year":"2006","journal-title":"J. Clin. Microbiol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1128\/JCM.02836-14","article-title":"Multisite clinical evaluation of a rapid test for Entamoeba histolytica in stool","volume":"53","author":"Verkerke","year":"2015","journal-title":"J. Clin. Microbiol."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Costa, D., Razakandrainibe, R., Valot, S., Vannier, M., Sautour, M., Basmaciyan, L., Gargala, G., Viller, V., Lemeteil, D., and Ballet, J.-J. (2020). Epidemiology of Cryptosporidiosis in France from 2017 to 2019. Microorganisms, 8.","DOI":"10.3390\/microorganisms8091358"}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/9\/11\/2325\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:28:13Z","timestamp":1760167693000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/9\/11\/2325"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,11,10]]},"references-count":38,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2021,11]]}},"alternative-id":["microorganisms9112325"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms9112325","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,11,10]]}}}