{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,3]],"date-time":"2026-10-03T23:54:34Z","timestamp":1791071674471,"version":"4.1.0"},"reference-count":28,"publisher":"American Society for Microbiology","issue":"9","license":[{"start":{"date-parts":[[2013,9,1]],"date-time":"2013-09-01T00:00:00Z","timestamp":1377993600000},"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":[[2013,9]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Molecular diagnosis of malaria offers many potential advantages over microscopy, including identification of malaria to the species level in an era with few experienced microscopists. We developed high-throughput multiplex 5\u2032 nuclease quantitative PCR (qPCR) assays, with the potential to support large studies, to specifically identify\n            <jats:named-content content-type=\"genus-species\">Plasmodium falciparum<\/jats:named-content>\n            ,\n            <jats:named-content content-type=\"genus-species\">P. vivax<\/jats:named-content>\n            ,\n            <jats:named-content content-type=\"genus-species\">P. ovale<\/jats:named-content>\n            ,\n            <jats:named-content content-type=\"genus-species\">P. malariae<\/jats:named-content>\n            , and\n            <jats:named-content content-type=\"genus-species\">P. knowlesi<\/jats:named-content>\n            . We compared qPCR to microscopy and confirmed discordant results with an alternative target PCR assay. The assays specifically detected 1 to 6 parasites\/\u03bcl of blood. The clinical sensitivities (95% confidence intervals [CIs]) of the 4-plex assay to detect microscopically confirmed malaria were 95.8% (88.3 to 99.1%) for\n            <jats:named-content content-type=\"genus-species\">P. falciparum<\/jats:named-content>\n            , 89.5% (75.2 to 97.1%) for\n            <jats:named-content content-type=\"genus-species\">P. vivax<\/jats:named-content>\n            , 94.1% (71.3 to 99.9%) for\n            <jats:named-content content-type=\"genus-species\">P. ovale<\/jats:named-content>\n            , and 100% (66.4 to 100%) for\n            <jats:named-content content-type=\"genus-species\">P. malariae<\/jats:named-content>\n            . The specificities (95% CIs) were 98.6% (92.4 to 100%) for\n            <jats:named-content content-type=\"genus-species\">P. falciparum<\/jats:named-content>\n            , 99% (84.8 to 100%) for\n            <jats:named-content content-type=\"genus-species\">P. vivax<\/jats:named-content>\n            , 98.4% (94.4 to 99.8%) for\n            <jats:named-content content-type=\"genus-species\">P. ovale<\/jats:named-content>\n            , and 99.3% (95.9 to 100%) for\n            <jats:named-content content-type=\"genus-species\">P. malariae<\/jats:named-content>\n            . The clinical specificity for samples without malaria was 100%. The clinical sensitivity of the 5-plex assay for confirmed\n            <jats:named-content content-type=\"genus-species\">P. knowlesi<\/jats:named-content>\n            malaria was 100% (95% CI, 69.2 to 100%), and the clinical specificity was 100% (95% CI, 87.2 to 100%). Coded retesting and testing with an alternative target PCR assay showed improved sensitivity and specificity of multiplex qPCR versus microscopy. Additionally, 91.7% (11\/12) of the samples with uncertain species by microscopy were identified to the species level identically by both our multiplex qPCR assay and the alternative target PCR assay, including 9\n            <jats:named-content content-type=\"genus-species\">P. falciparum<\/jats:named-content>\n            infections. Multiplex qPCR can rapidly and simultaneously identify all 5\n            <jats:named-content content-type=\"genus-species\">Plasmodium<\/jats:named-content>\n            species known to cause malaria in humans, and it offers an alternative or adjunct to microscopy for clinical diagnosis as well as a needed high-throughput tool for research.\n          <\/jats:p>","DOI":"10.1128\/jcm.00958-13","type":"journal-article","created":{"date-parts":[[2013,6,27]],"date-time":"2013-06-27T04:23:55Z","timestamp":1372307035000},"page":"2931-2938","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":38,"title":["Multiplex 5\u2032 Nuclease Quantitative Real-Time PCR for Clinical Diagnosis of Malaria and Species-Level Identification and Epidemiologic Evaluation of Malaria-Causing Parasites, Including Plasmodium knowlesi"],"prefix":"10.1128","volume":"51","author":[{"given":"Megan E.","family":"Reller","sequence":"first","affiliation":[{"name":"Division of Medical Microbiology, Department of Pathology, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wan Hsin","family":"Chen","sequence":"additional","affiliation":[{"name":"Division of Medical Microbiology, Department of Pathology, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Justin","family":"Dalton","sequence":"additional","affiliation":[{"name":"Division of Medical Microbiology, Department of Pathology, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marguerite A.","family":"Lichay","sequence":"additional","affiliation":[{"name":"Division of Medical Microbiology, Department of Pathology, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J. Stephen","family":"Dumler","sequence":"additional","affiliation":[{"name":"Division of Medical Microbiology, Department of Pathology, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","first-page":"42","volume-title":"World malaria report 2011, vol","author":"WHO Global Malaria Programme","year":"2011","unstructured":"WHO Global Malaria Programme. 2011. World malaria report 2011, vol 42. World Health Organization, Geneva, Switzerland."},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1093\/cid\/cis228"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0040633"},{"key":"e_1_3_2_5_2","volume-title":"Diagnostic methods in malaria","author":"Gilles H","year":"1993","unstructured":"GillesH. 1993. Diagnostic methods in malaria. 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