{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T05:44:54Z","timestamp":1790833494631,"version":"4.1.0"},"reference-count":25,"publisher":"American Society for Microbiology","issue":"6","license":[{"start":{"date-parts":[[2002,6,1]],"date-time":"2002-06-01T00:00:00Z","timestamp":1022889600000},"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":[[2002,6]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Detection and enumeration of\n            <jats:italic>Cryptosporidium parvum<\/jats:italic>\n            in both treated and untreated waters are important to facilitate prevention of future cryptosporidiosis incidents. Immunomagnetic separation (IMS)-fluorescent antibody (FA) detection and IMS-PCR detection efficiencies were evaluated in two natural waters seeded with nominal seed doses of 5, 10, and 15 oocysts. IMS-FA detected oocysts at concentrations at or below the three nominal oocyst seed doses, illustrating that IMS-FA is sensitive enough to detect low oocyst numbers. However, the species of the oocysts could not be determined with this technique. IMS-PCR, targeting the 18S rRNA gene in this study, yielded positive amplification for 17 of the 18 seeded water samples, and the amplicons were subjected to restriction fragment length polymorphism digestion and DNA sequencing for species identification. Interestingly, the two unseeded, natural water samples were also PCR positive; one amplicon was the same base pair size as the\n            <jats:italic>C. parvum<\/jats:italic>\n            amplicon, and the other amplicon was larger. These two amplified products were determined to be derived from DNA of\n            <jats:italic>Cryptosporidium muris<\/jats:italic>\n            and a dinoflagellate. These IMS-PCR results illustrate that (i) IMS-PCR is able to detect low oocyst numbers in natural waters, (ii) PCR amplification alone is not confirmatory for detection of target DNA when environmental samples are used, (ii) PCR primers, especially those designed against the rRNA gene region, need to be evaluated for specificity with organisms closely related to the target organism, and (iv) environmental amplicons should be subjected to appropriate species-specific confirmatory techniques.\n          <\/jats:p>","DOI":"10.1128\/aem.68.6.2991-2996.2002","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T10:09:29Z","timestamp":1027764569000},"page":"2991-2996","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":41,"title":["Immunomagnetic Separation (IMS)-Fluorescent Antibody Detection and IMS-PCR Detection of Seeded\n            <i>Cryptosporidium parvum<\/i>\n            Oocysts in Natural Waters and Their Limitations"],"prefix":"10.1128","volume":"68","author":[{"given":"Gregory D.","family":"Sturbaum","sequence":"first","affiliation":[{"name":"Department of Veterinary Science and Microbiology, University of Arizona, Tucson, Arizona 85721"},{"name":"CH Diagnostic &amp; Consulting Service, Inc., Loveland, Colorado 80537"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patricia T.","family":"Klonicki","sequence":"additional","affiliation":[{"name":"CH Diagnostic &amp; Consulting Service, Inc., Loveland, Colorado 80537"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marilyn M.","family":"Marshall","sequence":"additional","affiliation":[{"name":"Department of Veterinary Science and Microbiology, University of Arizona, Tucson, Arizona 85721"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"B. Helen","family":"Jost","sequence":"additional","affiliation":[{"name":"Department of Veterinary Science and Microbiology, University of Arizona, Tucson, Arizona 85721"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brec L.","family":"Clay","sequence":"additional","affiliation":[{"name":"CH Diagnostic &amp; Consulting Service, Inc., Loveland, Colorado 80537"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Charles R.","family":"Sterling","sequence":"additional","affiliation":[{"name":"Department of Veterinary Science and Microbiology, University of Arizona, Tucson, Arizona 85721"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"crossref","unstructured":"Altschul S. F. T. L. Madden A. A. Schaffer J. Zhang Z. Zhang W. Miller and D. J. Lipman. 1997. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. 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Lisle, and M. LeChevallier. 1997. Waterborne cryptosporidiosis: incidence, outbreaks, and treatment strategies, p. 93-109. In R. Fayer (ed.), Cryptosporidium and cryptosporidiosis. CRC Press, Inc., New York, N.Y.","journal-title":"Cryptosporidium and cryptosporidiosis"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0169-4758(97)01150-2"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1099\/0022-1317-49-3-207"},{"key":"e_1_3_2_25_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.67.6.2665-2668.2001"},{"key":"e_1_3_2_26_2","unstructured":"United States Environmental Protection Agency. 1999. Method 1623: Cryptosporidium and Giardia in water by filtration\/IMS\/FA. Publication EPA-821-R-99-006. 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