{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T18:31:49Z","timestamp":1792175509290,"version":"4.4.0"},"reference-count":49,"publisher":"American Society for Microbiology","issue":"8","license":[{"start":{"date-parts":[[2012,8,1]],"date-time":"2012-08-01T00:00:00Z","timestamp":1343779200000},"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":[[2012,8]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            A single-step multiplex PCR (here referred to as a duplex PCR) has been developed for simultaneous detection and diagnosis of\n            <jats:named-content content-type=\"genus-species\">Fasciola hepatica<\/jats:named-content>\n            and\n            <jats:named-content content-type=\"genus-species\">F. gigantica<\/jats:named-content>\n            . These species overlap in distribution in many countries of North and East Africa and Central and Southeast Asia and are similar in egg morphology, making identification from fecal samples difficult. Based on a comparative alignment of mitochondrial DNA (mtDNA) spanning the region of\n            <jats:italic>cox1-trnT-rrnL<\/jats:italic>\n            , two species-specific forward primers were designed, FHF (for\n            <jats:named-content content-type=\"genus-species\">F. hepatica<\/jats:named-content>\n            ) and FGF (for\n            <jats:named-content content-type=\"genus-species\">F. gigantica<\/jats:named-content>\n            ), and a single reverse primer, FHGR (common for both species). Conventional PCR followed by sequencing was applied using species-specific primer pairs to verify the specificity of primers and the identity of\n            <jats:named-content content-type=\"genus-species\">Fasciola<\/jats:named-content>\n            DNA templates. Duplex PCR (using three primers) was used for testing with the DNA extracted from adult worms, miracidia, and eggs, producing amplicons of 1,031 bp for\n            <jats:named-content content-type=\"genus-species\">F. hepatica<\/jats:named-content>\n            and 615 bp for\n            <jats:named-content content-type=\"genus-species\">F. gigantica<\/jats:named-content>\n            . The duplex PCR failed to amplify from DNA of other common liver and intestinal trematodes, including two opisthorchiids, three heterophyids, an echinostomid, another fasciolid, and a taeniid cestode. The sensitivity assay showed that the duplex PCR limit of detection for each\n            <jats:named-content content-type=\"genus-species\">Fasciola<\/jats:named-content>\n            species was between 0.012 ng and 0.006 ng DNA. Evaluation using DNA templates from 32\n            <jats:named-content content-type=\"genus-species\">Fasciola<\/jats:named-content>\n            samples (28 adults and 4 eggs) and from 25 field-collected stools of ruminants and humans revealed specific bands of the correct size and the presence of\n            <jats:named-content content-type=\"genus-species\">Fasciola<\/jats:named-content>\n            species. This novel mtDNA duplex PCR is a sensitive and fast tool for accurate identification of\n            <jats:named-content content-type=\"genus-species\">Fasciola<\/jats:named-content>\n            species in areas of distributional and zonal overlap.\n          <\/jats:p>","DOI":"10.1128\/jcm.00662-12","type":"journal-article","created":{"date-parts":[[2012,6,13]],"date-time":"2012-06-13T04:57:14Z","timestamp":1339563434000},"page":"2720-2726","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":39,"title":["Development and Evaluation of a Single-Step Duplex PCR for Simultaneous Detection of Fasciola hepatica and Fasciola gigantica (Family Fasciolidae, Class Trematoda, Phylum Platyhelminthes)"],"prefix":"10.1128","volume":"50","author":[{"given":"Thanh Hoa","family":"Le","sequence":"first","affiliation":[{"name":"Department of Immunology, Institute of Biotechnology, Vietnam Academy of Science and Technology, Cau Giay, Hanoi, Vietnam"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Khue Thi","family":"Nguyen","sequence":"additional","affiliation":[{"name":"Department of Immunology, Institute of Biotechnology, Vietnam Academy of Science and Technology, Cau Giay, Hanoi, Vietnam"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nga Thi Bich","family":"Nguyen","sequence":"additional","affiliation":[{"name":"Department of Immunology, Institute of Biotechnology, Vietnam Academy of Science and Technology, Cau Giay, Hanoi, Vietnam"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huong Thi Thanh","family":"Doan","sequence":"additional","affiliation":[{"name":"Department of Immunology, Institute of Biotechnology, Vietnam Academy of Science and Technology, Cau Giay, Hanoi, Vietnam"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuyen Thi Kim","family":"Le","sequence":"additional","affiliation":[{"name":"Department of Immunology, Institute of Biotechnology, Vietnam Academy of Science and Technology, Cau Giay, Hanoi, Vietnam"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chau Thi Minh","family":"Hoang","sequence":"additional","affiliation":[{"name":"Department of Immunology, Institute of Biotechnology, Vietnam Academy of Science and Technology, Cau Giay, Hanoi, Vietnam"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nguyen Van","family":"De","sequence":"additional","affiliation":[{"name":"Department of Parasitology, Hanoi Medical University, Dong Da, Hanoi, Vietnam"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1186\/1756-3305-4-101"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1179\/136485910X12607012373713"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetpar.2010.09.005"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetpar.2011.03.057"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetpar.2011.01.059"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00436-007-0870-7"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.parint.2010.09.003"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.exppara.2011.03.011"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.parint.2006.06.003"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetpar.2010.12.020"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00436-008-1086-1"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetpar.2008.01.035"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1186\/1756-3305-4-176"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.3390\/genes1010102"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.diagmicrobio.2011.03.016"},{"key":"e_1_3_2_17_2","volume-title":"Promega Notes.","author":"Frackman S","year":"1998","unstructured":"FrackmanS KobsG SimpsonD StortsD. 1998. 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