{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,24]],"date-time":"2025-09-24T10:30:31Z","timestamp":1758709831694},"reference-count":24,"publisher":"American Society for Microbiology","issue":"2","license":[{"start":{"date-parts":[[2005,2,1]],"date-time":"2005-02-01T00:00:00Z","timestamp":1107216000000},"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":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2005,2]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            <jats:italic>Cryptosporidium parvum<\/jats:italic>\n            and\n            <jats:italic>C. hominis<\/jats:italic>\n            have been the cause of large and serious outbreaks of waterborne cryptosporidiosis. A specific and sensitive recovery-detection method is required for control of this pathogen in drinking water. In the present study, nested PCR-restriction fragment length polymorphism (RFLP), which targets the divergent\n            <jats:italic>Cpgp40\/15<\/jats:italic>\n            gene, was developed. This nested PCR detected only the gene derived from\n            <jats:italic>C. parvum<\/jats:italic>\n            and\n            <jats:italic>C. hominis<\/jats:italic>\n            strains, and RFLP was able to discriminate between the PCR products from\n            <jats:italic>C. parvum<\/jats:italic>\n            and\n            <jats:italic>C. hominis<\/jats:italic>\n            . To evaluate the sensitivity of nested PCR,\n            <jats:italic>C. parvum<\/jats:italic>\n            oocysts inoculated in water samples of two different turbidities were recovered by immunomagnetic separation (IMS) and detected by nested PCR and fluorescent antibody assay (FA). Genetic detection by nested PCR and oocyst number confirmed by FA were compared, and the results suggested that detection by nested PCR depends on the confirmed oocyst number and that nested PCR in combination with IMS has the ability to detect a single oocyst in a water sample. We applied an agitation procedure with river water solids to which oocysts were added to evaluate the recovery and detection by the procedure in environmental samples and found some decrease in the rate of detection by IMS.\n          <\/jats:p>","DOI":"10.1128\/aem.71.2.898-903.2005","type":"journal-article","created":{"date-parts":[[2005,2,3]],"date-time":"2005-02-03T18:24:28Z","timestamp":1107455068000},"page":"898-903","update-policy":"http:\/\/dx.doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":17,"title":["Detection and Discrimination of\n            <i>Cryptosporidium parvum<\/i>\n            and\n            <i>C. hominis<\/i>\n            in Water Samples by Immunomagnetic Separation-PCR"],"prefix":"10.1128","volume":"71","author":[{"given":"Yoshitsugu","family":"Ochiai","sequence":"first","affiliation":[{"name":"Division of Water Environment, Tokyo Metropolitan Institute of Public Health, Shinjuku-ku, Tokyo, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chieko","family":"Takada","sequence":"additional","affiliation":[{"name":"Division of Water Environment, Tokyo Metropolitan Institute of Public Health, Shinjuku-ku, Tokyo, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mitsugu","family":"Hosaka","sequence":"additional","affiliation":[{"name":"Division of Water Environment, Tokyo Metropolitan Institute of Public Health, Shinjuku-ku, Tokyo, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.64.11.4495-4499.1998"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM199503303321304"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.69.4.1898-1903.2003"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0166-0934(96)02123-4"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.61.11.3849-3855.1995"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.56.5.1423-1428.1990"},{"key":"e_1_3_2_8_2","first-page":"103","volume":"44","year":"1995","unstructured":"Lisle, J. 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Cold Spring Harbor Laboratory Cold Spring Harbor N.Y."},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0167-7012(01)00250-0"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.38.3.1180-1183.2000"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1550-7408.2002.tb00224.x"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1086\/315033"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid0304.970423"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1086\/342296"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.2166\/wst.2000.0121"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.68.7.4117-4134.2000"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.68.6.2991-2996.2002"},{"key":"e_1_3_2_21_2","unstructured":"U.S. Environmental Protection Agency. 1999. Method 1623: Cryptosporidium and Giardia in water by filtration\/IMS\/FA. Publication EPA-821-R-99-006. U.S. Environmental Protection Agency Washington D.C."},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1086\/318090"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.65.4.1578-1583.1999"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.17.1.72-97.2004"},{"key":"e_1_3_2_25_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.67.3.1097-1101.2001"}],"container-title":["Applied and Environmental Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AEM.71.2.898-903.2005","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AEM.71.2.898-903.2005","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,2,23]],"date-time":"2022-02-23T15:14:22Z","timestamp":1645629262000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/AEM.71.2.898-903.2005"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2005,2]]},"references-count":24,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2005,2]]}},"alternative-id":["10.1128\/AEM.71.2.898-903.2005"],"URL":"https:\/\/doi.org\/10.1128\/aem.71.2.898-903.2005","relation":{},"ISSN":["0099-2240","1098-5336"],"issn-type":[{"value":"0099-2240","type":"print"},{"value":"1098-5336","type":"electronic"}],"subject":[],"published":{"date-parts":[[2005,2]]},"assertion":[{"value":"2004-05-16","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2004-09-21","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2005-02-01","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}