{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T10:39:40Z","timestamp":1791196780355,"version":"4.1.0"},"reference-count":37,"publisher":"American Society for Microbiology","issue":"8","license":[{"start":{"date-parts":[[2013,8,1]],"date-time":"2013-08-01T00:00:00Z","timestamp":1375315200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Clin Microbiol"],"published-print":{"date-parts":[[2013,8]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            <jats:named-content content-type=\"genus-species\">Cryptosporidium<\/jats:named-content>\n            is a protozoan parasite responsible for gastroenteritis, especially in immunocompromised patients. Laboratory diagnosis of cryptosporidiosis relies on microscopy, antigen detection, and nucleic acid detection and analysis. Among the numerous molecular targets available, the 18S rRNA gene displays the best sensitivity and sequence variations between species and can be used for molecular typing assays. This paper presents a new real-time PCR assay for the detection and quantification of all\n            <jats:named-content content-type=\"genus-species\">Cryptosporidium<\/jats:named-content>\n            species associated with the identification of\n            <jats:named-content content-type=\"genus-species\">Cryptosporidium hominis<\/jats:named-content>\n            and\n            <jats:named-content content-type=\"genus-species\">Cryptosporidium parvum<\/jats:named-content>\n            . The sensitivity and specificity of this new PCR assay were assessed on a multicentric basis, using well-characterized\n            <jats:named-content content-type=\"genus-species\">Cryptosporidium<\/jats:named-content>\n            -positive and -negative human stool samples, and the efficiencies of nine extraction methods were comparatively assessed using\n            <jats:named-content content-type=\"genus-species\">Cryptosporidium<\/jats:named-content>\n            -seeded stool samples and phosphate-buffered saline samples. A comparison of extraction yields showed that the most efficient extraction method was the Boom technique in association with mechanical grinding, and column extraction showed higher binding capacity than extraction methods based on magnetic silica. Our PCR assay was able to quantify at least 300 oocysts per gram of stool. Satisfactory reproducibility between laboratories was observed. The two main species causing human disease,\n            <jats:named-content content-type=\"genus-species\">Cryptosporidium hominis<\/jats:named-content>\n            and\n            <jats:named-content content-type=\"genus-species\">Cryptosporidium parvum<\/jats:named-content>\n            , were identified using a duplex real-time PCR assay with specific TaqMan minor-groove-binding ligand (MGB) probes for the same amplicon. To conclude, this one-step quantitative PCR is well suited to the routine diagnosis of cryptosporidiosis since practical conditions, including DNA extraction, quantification using well-defined standards, and identification of the two main species infecting humans, have been positively assessed.\n          <\/jats:p>","DOI":"10.1128\/jcm.03458-12","type":"journal-article","created":{"date-parts":[[2013,5,30]],"date-time":"2013-05-30T04:48:07Z","timestamp":1369889287000},"page":"2556-2563","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":79,"title":["Multicentric Evaluation of a New Real-Time PCR Assay for Quantification of Cryptosporidium spp. and Identification of Cryptosporidium parvum and Cryptosporidium hominis"],"prefix":"10.1128","volume":"51","author":[{"given":"C.","family":"Mary","sequence":"first","affiliation":[{"name":"Aix-Marseille Universit\u00e9, Facult\u00e9 de M\u00e9decine, UMR MD3, and APHM, H\u00f4pital de la Timone, Laboratoire de Parasitologie-Mycologie, Marseille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"E.","family":"Chapey","sequence":"additional","affiliation":[{"name":"Laboratoire de Parasitologie-Mycologie, Hospices Civils de Lyon, Universit\u00e9 Lyon 1, Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"E.","family":"Dutoit","sequence":"additional","affiliation":[{"name":"Laboratoire de Parasitologie-Mycologie, Centre Hospitalo-Universitaire de Lille, Lille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"K.","family":"Guyot","sequence":"additional","affiliation":[{"name":"Biologie et Diversit\u00e9 des Pathog\u00e8nes Eucaryotes \u00c9mergents, Centre d'Infection et d'Immunit\u00e9 de Lille, Institut Pasteur de Lille, INSERM U1019, CNRS UMR 8204, Universit\u00e9 Lille-Nord-de-France, Lille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"L.","family":"Hasseine","sequence":"additional","affiliation":[{"name":"Laboratoire de Parasitologie-Mycologie, Centre Hospitalo-Universitaire de Nice, Nice, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"F.","family":"Jeddi","sequence":"additional","affiliation":[{"name":"Aix-Marseille Universit\u00e9, Facult\u00e9 de M\u00e9decine, UMR MD3, and APHM, H\u00f4pital de la Timone, Laboratoire de Parasitologie-Mycologie, Marseille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J.","family":"Menotti","sequence":"additional","affiliation":[{"name":"Laboratoire de Parasitologie-Mycologie, H\u00f4pital Saint-Louis, Assistance Publique-H\u00f4pitaux de Paris and Universit\u00e9 Denis Diderot, Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C.","family":"Paraud","sequence":"additional","affiliation":[{"name":"Agence Nationale de S\u00e9curit\u00e9 Sanitaire de l'Alimentation, de l'Environnement, et du Travail, Niort, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C.","family":"Pomares","sequence":"additional","affiliation":[{"name":"Laboratoire de Parasitologie-Mycologie, Centre Hospitalo-Universitaire de Nice, Nice, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M.","family":"Rabodonirina","sequence":"additional","affiliation":[{"name":"Laboratoire de Parasitologie-Mycologie, Hospices Civils de Lyon, Universit\u00e9 Lyon 1, Lyon, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A.","family":"Rieux","sequence":"additional","affiliation":[{"name":"Agence Nationale de S\u00e9curit\u00e9 Sanitaire de l'Alimentation, de l'Environnement, et du Travail, Niort, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"F.","family":"Derouin","sequence":"additional","affiliation":[{"name":"Laboratoire de Parasitologie-Mycologie, H\u00f4pital Saint-Louis, Assistance Publique-H\u00f4pitaux de Paris and Universit\u00e9 Denis Diderot, Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"name":"for the ANOFEL Cryptosporidium National Network","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.exppara.2009.02.003"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1586\/14787210.6.3.337"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1017\/S0950268811000860"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1017\/S0031182009990436"},{"key":"e_1_3_2_6_2","doi-asserted-by":"crossref","unstructured":"ANOFEL Cryptosporidium National Network. 2010. 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