{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T07:07:07Z","timestamp":1781852827967,"version":"3.54.5"},"reference-count":41,"publisher":"American Society for Microbiology","issue":"6","license":[{"start":{"date-parts":[[2000,6,1]],"date-time":"2000-06-01T00:00:00Z","timestamp":959817600000},"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":[[2000,6]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Within the last several years, the protozoan parasites\n            <jats:italic>Cyclospora cayetanensis<\/jats:italic>\n            ,\n            <jats:italic>Cryptosporidium parvum<\/jats:italic>\n            , and microsporidia have become recognized as important, rapidly emerging human pathogens in immunocompromised and immunocompetent individuals. Since the early 1990s, many of the reported outbreaks of enteric illness caused by these microorganisms have been attributed to food- and water-borne contamination. Many inherent obstacles affect the success of current surveillance and detection methods used to monitor and control levels of contamination by these pathogens. Unlike methods that incorporate preenrichment for easier and unambiguous identification of bacterial pathogens, similar methods for the detection of parasitic protozoa either are not currently available or cannot be performed in a timely manner. We have developed an extraction-free, filter-based protocol to prepare DNA templates for use in PCR to identify\n            <jats:italic>C. cayetanensis<\/jats:italic>\n            and\n            <jats:italic>C. parvum<\/jats:italic>\n            oocysts and microsporidia spores. This method requires only minimal preparation to partially purify and concentrate isolates prior to filter application. DNA template preparation is rapid, efficient, and reproducible. As few as 3 to 10 parasites could be detected by PCR from direct application to the filters. In studies, as few 10 to 50\n            <jats:italic>Encephalitozoon intestinalis<\/jats:italic>\n            spores could be detected when seeded in a 100-\u03bcl stool sample and 10 to 30\n            <jats:italic>C. cayetanensis<\/jats:italic>\n            oocysts could be detected per 100 g of fresh raspberries. This protocol can easily be adapted to detect parasites from a wide variety of food, clinical, and environmental samples and can be used in multiplex PCR applications.\n          <\/jats:p>","DOI":"10.1128\/jcm.38.6.2271-2277.2000","type":"journal-article","created":{"date-parts":[[2020,3,9]],"date-time":"2020-03-09T19:47:22Z","timestamp":1583783242000},"page":"2271-2277","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":115,"title":["Extraction-Free, Filter-Based Template Preparation for Rapid and Sensitive PCR Detection of Pathogenic Parasitic Protozoa"],"prefix":"10.1128","volume":"38","author":[{"given":"Palmer A.","family":"Orlandi","sequence":"first","affiliation":[{"name":"<!--label omitted: 1-->Division Virulence Assessment, Center for Food Safety and Applied Nutrition, Food and Drug Administration, Washington, D.C. 20204"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Keith A.","family":"Lampel","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Division Virulence Assessment, Center for Food Safety and Applied Nutrition, Food and Drug Administration, Washington, D.C. 20204"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_1_2_2","first-page":"426","article-title":"Automated DNA purification and amplification from blood-stained cards using a robotic workstation","volume":"19","author":"Belgrader P.","year":"1995","unstructured":"Belgrader P. Del Rio S. A. Turner K. A. Marino M. A. Weaver K. R. Williams P. E. Automated DNA purification and amplification from blood-stained cards using a robotic workstation.Biotechniques191995426432","journal-title":"Biotechniques"},{"key":"e_1_3_1_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.33.10.2592-2595.1995"},{"key":"e_1_3_1_4_2","unstructured":"Burgoyne L. A. March 1996. Solid medium and method for DNA storage. U.S. patent 5 496 562."},{"key":"e_1_3_1_5_2","first-page":"325","article-title":"Outbreaks of diarrheal illness associated with cyanobacteria (blue-green algae)-like bodies\u2014Chicago and Nepal, 1989 and 1990","volume":"40","author":"Centers for Disease Control","year":"1991","unstructured":"Centers for Disease Control Outbreaks of diarrheal illness associated with cyanobacteria (blue-green algae)-like bodies\u2014Chicago and Nepal, 1989 and 1990.Morbid. Mortal. Wkly. Rep.401991325327","journal-title":"Morbid. Mortal. Wkly. Rep."},{"key":"e_1_3_1_6_2","first-page":"4","article-title":"Outbreaks of Escherichia coli O175:H7 infection and cryptosporidiosis associated with drinking unpasteurized apple cider\u2014Connecticut and New York, October 1996","volume":"46","author":"Centers for Disease Control","year":"1997","unstructured":"Centers for Disease Control Outbreaks of Escherichia coli O175:H7 infection and cryptosporidiosis associated with drinking unpasteurized apple cider\u2014Connecticut and New York, October 1996.Morbid. Mortal. Wkly. Rep.46199748","journal-title":"Morbid. Mortal. Wkly. Rep."},{"key":"e_1_3_1_7_2","first-page":"521","article-title":"Update: outbreaks of cyclosporiasis\u2014United States and Canada","volume":"46","author":"Centers for Disease Control","year":"1997","unstructured":"Centers for Disease Control Update: outbreaks of cyclosporiasis\u2014United States and Canada.Morbid. Mortal. Wkly. Rep.461997521523","journal-title":"Morbid. Mortal. Wkly. Rep."},{"key":"e_1_3_1_8_2","first-page":"689","article-title":"Outbreak of cyclosporiasis\u2014Northern Virginia-Washington, D.C.-Baltimore, Maryland, metropolitan area, 1997","volume":"46","author":"Centers for Disease Control","year":"1997","unstructured":"Centers for Disease Control Outbreak of cyclosporiasis\u2014Northern Virginia-Washington, D.C.-Baltimore, Maryland, metropolitan area, 1997.Morbid. Mortal. Wkly. Rep.461997689691","journal-title":"Morbid. Mortal. Wkly. Rep."},{"key":"e_1_3_1_9_2","first-page":"806","article-title":"Update: outbreak of cyclosporiasis\u2014Ontario, Canada, May 1998","volume":"47","author":"Centers for Disease Control","year":"1998","unstructured":"Centers for Disease Control Update: outbreak of cyclosporiasis\u2014Ontario, Canada, May 1998.Morbid. Mortal. Wkly. Rep.471998806809","journal-title":"Morbid. Mortal. Wkly. Rep."},{"key":"e_1_3_1_10_2","doi-asserted-by":"crossref","first-page":"S143","DOI":"10.1017\/S0031182099004904","article-title":"Emerging pathogens: Isospora, Cyclospora, and Microsporidia","volume":"117","author":"Curry A.","year":"1998","unstructured":"Curry A. Smith H. V. Emerging pathogens: Isospora, Cyclospora, and Microsporidia.Parasitology117 (Suppl)1998S143S159","journal-title":"Parasitology"},{"key":"e_1_3_1_11_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.34.4.986-987.1996"},{"key":"e_1_3_1_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1084-8592(97)80010-0"},{"key":"e_1_3_1_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1084-8592(99)80050-2"},{"key":"e_1_3_1_14_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/23.13.2567"},{"key":"e_1_3_1_15_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.35.7.1862-1866.1997"},{"key":"e_1_3_1_16_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.64.9.3332-3335.1998"},{"key":"e_1_3_1_17_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid0203.960304"},{"key":"e_1_3_1_18_2","doi-asserted-by":"publisher","DOI":"10.3928\/0090-4481-19960901-04"},{"key":"e_1_3_1_19_2","doi-asserted-by":"publisher","DOI":"10.7326\/0003-4819-124-4-199602150-00008"},{"key":"e_1_3_1_20_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM198905253202103"},{"key":"e_1_3_1_21_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM199705293362202"},{"key":"e_1_3_1_22_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(95)90868-4"},{"key":"e_1_3_1_23_2","doi-asserted-by":"publisher","DOI":"10.4315\/0362-028X-61.11.1497"},{"key":"e_1_3_1_24_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.61.11.3849-3855.1995"},{"key":"e_1_3_1_25_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF02760775"},{"key":"e_1_3_1_26_2","doi-asserted-by":"publisher","DOI":"10.1016\/0168-1605(96)00977-4"},{"key":"e_1_3_1_27_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.64.11.4234-4237.1998"},{"key":"e_1_3_1_28_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM199407213310304"},{"key":"e_1_3_1_29_2","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(91)90292-2"},{"key":"e_1_3_1_30_2","doi-asserted-by":"publisher","DOI":"10.1001\/jama.1994.03520200048034"},{"key":"e_1_3_1_31_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM199305063281804"},{"key":"e_1_3_1_32_2","doi-asserted-by":"publisher","DOI":"10.4269\/ajtmh.1997.57.683"},{"key":"e_1_3_1_33_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(94)90716-1"},{"key":"e_1_3_1_34_2","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/173.2.440"},{"key":"e_1_3_1_35_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.36.6.1814-1818.1998"},{"key":"e_1_3_1_36_2","doi-asserted-by":"publisher","DOI":"10.1006\/abio.1997.2031"},{"key":"e_1_3_1_37_2","doi-asserted-by":"publisher","DOI":"10.1007\/s002530051057"},{"key":"e_1_3_1_38_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.64.6.2284-2286.1998"},{"key":"e_1_3_1_39_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.32.11.2760-2768.1994"},{"key":"e_1_3_1_40_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.33.4.930-936.1995"},{"key":"e_1_3_1_41_2","doi-asserted-by":"publisher","DOI":"10.1093\/clinids\/18.4.620"},{"key":"e_1_3_1_42_2","unstructured":"Yoder\nK. E.\nSethabutr\nO.\nRelman\nD. A.\nPCR-based detection of the intestinal pathogen Cyclospora PCR protocols for emerging infectious diseases a supplement to diagnostic molecular microbiology: principles and applications.\nPersing\nD. H.\n1996\n169\n176\nASM Press\nWashington D.C."}],"container-title":["Journal of Clinical Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/JCM.38.6.2271-2277.2000","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/JCM.38.6.2271-2277.2000","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,3,5]],"date-time":"2022-03-05T00:41:26Z","timestamp":1646440886000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/JCM.38.6.2271-2277.2000"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2000,6]]},"references-count":41,"aliases":["10.1128\/.38.6.2271-2277.2000"],"journal-issue":{"issue":"6","published-print":{"date-parts":[[2000,6]]}},"alternative-id":["10.1128\/JCM.38.6.2271-2277.2000"],"URL":"https:\/\/doi.org\/10.1128\/jcm.38.6.2271-2277.2000","relation":{},"ISSN":["0095-1137","1098-660X"],"issn-type":[{"value":"0095-1137","type":"print"},{"value":"1098-660X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2000,6]]},"assertion":[{"value":"2000-01-05","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2000-03-08","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2000-06-01","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}