{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,9]],"date-time":"2026-07-09T13:00:53Z","timestamp":1783602053378,"version":"3.55.0"},"reference-count":32,"publisher":"American Society for Microbiology","issue":"1","license":[{"start":{"date-parts":[[2011,1,1]],"date-time":"2011-01-01T00:00:00Z","timestamp":1293840000000},"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":[[2011,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            The aim of this study was to describe the first development and evaluation of a multiplex tandem PCR (MT-PCR) assay for the detection and identification of 4 common pathogenic protozoan parasites,\n            <jats:italic>Cryptosporidium<\/jats:italic>\n            spp.,\n            <jats:italic>Dientamoeba fragilis<\/jats:italic>\n            ,\n            <jats:italic>Entamoeba histolytica<\/jats:italic>\n            , and\n            <jats:italic>Giardia intestinalis<\/jats:italic>\n            , from human clinical samples. A total of 472 fecal samples submitted to the Department of Microbiology at St. Vincent's Hospital were included in the study. The MT-PCR assay was compared to four real-time PCR (RT-PCR) assays and microscopy by a traditional modified iron hematoxylin stain. The MT-PCR detected 28\n            <jats:italic>G. intestinalis<\/jats:italic>\n            , 26\n            <jats:italic>D. fragilis<\/jats:italic>\n            , 11\n            <jats:italic>E. histolytica<\/jats:italic>\n            , and 9\n            <jats:italic>Cryptosporidium<\/jats:italic>\n            sp. isolates. Detection and identification of the fecal protozoa by MT-PCR demonstrated 100% correlation with the RT-PCR results, and compared to RT-PCR, MT-PCR exhibited 100% sensitivity and specificity, while traditional microscopy of stained fixed fecal smears exhibited sensitivities and specificities of 56% and 100% for\n            <jats:italic>Cryptosporidium<\/jats:italic>\n            spp., 38% and 99% for\n            <jats:italic>D. fragilis<\/jats:italic>\n            , 47% and 97% for\n            <jats:italic>E. histolytica<\/jats:italic>\n            , and 50% and 100% for\n            <jats:italic>G. intestinalis<\/jats:italic>\n            . No cross-reactivity was detected in 100 stool samples containing various other bacterial, viral, and protozoan species. The MT-PCR assay was able to provide rapid, sensitive, and specific simultaneous detection and identification of the four most important diarrhea-causing protozoan parasites that infect humans. This study also highlights the lack of sensitivity demonstrated by microscopy, and thus, molecular methods such as MT-PCR must be considered the diagnostic methods of choice for enteric protozoan parasites.\n          <\/jats:p>","DOI":"10.1128\/jcm.01796-10","type":"journal-article","created":{"date-parts":[[2010,11,4]],"date-time":"2010-11-04T02:27:09Z","timestamp":1288837629000},"page":"257-262","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":116,"title":["Evaluation of Multiplex Tandem Real-Time PCR for Detection of\n            <i>Cryptosporidium<\/i>\n            spp.,\n            <i>Dientamoeba fragilis<\/i>\n            ,\n            <i>Entamoeba histolytica<\/i>\n            , and\n            <i>Giardia intestinalis<\/i>\n            in Clinical Stool Samples"],"prefix":"10.1128","volume":"49","author":[{"given":"D.","family":"Stark","sequence":"first","affiliation":[{"name":"Division of Microbiology, SydPath, St. Vincent's Hospital, Sydney, Australia"},{"name":"University of Technology Sydney, Department of Medical and Molecular Biosciences, Broadway, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S. E.","family":"Al-Qassab","sequence":"additional","affiliation":[{"name":"University of Technology Sydney, Department of Medical and Molecular Biosciences, Broadway, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J. L. N.","family":"Barratt","sequence":"additional","affiliation":[{"name":"Division of Microbiology, SydPath, St. Vincent's Hospital, Sydney, Australia"},{"name":"University of Technology Sydney, Department of Medical and Molecular Biosciences, Broadway, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"K.","family":"Stanley","sequence":"additional","affiliation":[{"name":"AusDiagnostics Pty. Ltd., 3\/36 O'Riordan St., Alexandria, New South Wales, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"T.","family":"Roberts","sequence":"additional","affiliation":[{"name":"Division of Microbiology, SydPath, St. Vincent's Hospital, Sydney, Australia"},{"name":"University of Technology Sydney, Department of Medical and Molecular Biosciences, Broadway, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"D.","family":"Marriott","sequence":"additional","affiliation":[{"name":"Division of Microbiology, SydPath, St. Vincent's Hospital, Sydney, Australia"},{"name":"University of Technology Sydney, Department of Medical and Molecular Biosciences, Broadway, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J.","family":"Harkness","sequence":"additional","affiliation":[{"name":"Division of Microbiology, SydPath, St. Vincent's Hospital, Sydney, Australia"},{"name":"University of Technology Sydney, Department of Medical and Molecular Biosciences, Broadway, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J. 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