{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T04:42:11Z","timestamp":1788410531874,"version":"build-2803163510"},"reference-count":25,"publisher":"American Society of Tropical Medicine and Hygiene","issue":"5","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2018,11,7]]},"abstract":"<jats:p>Hookworm disease caused by <jats:italic>Necator americanus<\/jats:italic>, <jats:italic>Ancylostoma duodenale<\/jats:italic>, and <jats:italic>Ancylostoma ceylanicum<\/jats:italic> affects half a billion people worldwide. The prevalence and intensity of infection of individual hookworm species are vital for assessing morbidity and generating targeted intervention programs for their control. The present study aims to evaluate a multiplex real-time quantitative PCR (qPCR) assay to determine the prevalence and egg intensity of all three hookworm species and compare this with standard microscopy and published genus-based conventional and real-time multiplex qPCRs. Performance of the diagnostic assays was evaluated using DNA extracted from 192 fecal samples collected as part of a soil-transmitted helminth (STH) survey in northern Cambodia. The prevalence of hookworms as detected by the multiplex hookworm qPCR of 84\/192 (43.8%) was significantly higher than that using microscopy of 49\/192 (25.5%). The hookworm multiplex qPCR showed very good agreement for the detection of both <jats:italic>N. americanus<\/jats:italic> (Kappa 0.943) and <jats:italic>Ancylostoma<\/jats:italic> spp. (Kappa 0.936) with a multiplex STH qPCR. A strong and moderate quantitative correlation between cycle threshold and eggs per gram (EPG) feces was obtained for the hookworm qPCR for seeded DNA egg extracts (<jats:italic>R<\/jats:italic><jats:sup>2<\/jats:sup> \u2265 0.9004) and naturally egg-infected individuals (<jats:italic>R<\/jats:italic><jats:sup>2<\/jats:sup> = 0.6848), respectively. The newly developed hookworm quantitative multiplex qPCR has the potential for application in anthelmintic efficacy trials and for monitoring the success of mass deworming programs targeting individual species of anthroponotic and zoonotic hookworms.<\/jats:p>","DOI":"10.4269\/ajtmh.18-0276","type":"journal-article","created":{"date-parts":[[2018,9,18]],"date-time":"2018-09-18T07:01:10Z","timestamp":1537254070000},"page":"1186-1193","source":"Crossref","is-referenced-by-count":47,"title":["Development and Evaluation of a Multiplex Quantitative Real-Time Polymerase Chain Reaction for Hookworm Species in Human Stool"],"prefix":"10.4269","volume":"99","author":[{"given":"Sze Fui","family":"Hii","sequence":"first","affiliation":[{"name":"Faculty of Veterinary and Agricultural Sciences, University of Melbourne, Parkville, Australia;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dammika","family":"Senevirathna","sequence":"additional","affiliation":[{"name":"Faculty of Veterinary and Agricultural Sciences, University of Melbourne, Parkville, Australia;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stacey","family":"Llewellyn","sequence":"additional","affiliation":[{"name":"Clinical Tropical Medicine Laboratory, QIMR Berghofer Medical Research Institute, Herston, Australia;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tawin","family":"Inpankaew","sequence":"additional","affiliation":[{"name":"Department of Parasitology, Faculty of Veterinary Medicine, Kasetsart University, Bangkok, Thailand;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Odermatt","sequence":"additional","affiliation":[{"name":"Department of Epidemiology and Public Health, Swiss Tropical and Public Health Institute, Basel, Switzerland;"},{"name":"University of Basel, Basel, Switzerland;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Virak","family":"Khieu","sequence":"additional","affiliation":[{"name":"National Centre for Parasitology, Entomology, and Malaria Control, Phnom Penh, Cambodia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sinoun","family":"Muth","sequence":"additional","affiliation":[{"name":"National Centre for Parasitology, Entomology, and Malaria Control, Phnom Penh, Cambodia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"James","family":"McCarthy","sequence":"additional","affiliation":[{"name":"Clinical Tropical Medicine Laboratory, QIMR Berghofer Medical Research Institute, Herston, Australia;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rebecca J.","family":"Traub","sequence":"additional","affiliation":[{"name":"Faculty of Veterinary and Agricultural Sciences, University of Melbourne, Parkville, Australia;"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"2795","reference":[{"key":"b1","series-title":"Lancet","first-page":"2287","article-title":"Global, regional, and national comparative risk assessment of 79 behavioural, environmental and occupational, and metabolic risks or clusters of risks in 188 countries, 1990\u20132013: a systematic analysis for the Global Burden of Disease Study 2013","volume":"386","author":"Forouzanfar","year":"2015"},{"key":"b2","series-title":"Obstet Gynecol","first-page":"155","article-title":"Helminth infections: a new global women\u2019s health agenda","volume":"123","author":"Hotez","year":"2014"},{"key":"b3","series-title":"Int J Parasitol","first-page":"1009","article-title":"Ancylostoma ceylanicum, a re-emerging but neglected parasitic zoonosis","volume":"43","author":"Traub","year":"2013"},{"key":"b4","series-title":"Emerg Infect Dis","first-page":"976","article-title":"High prevalence of Ancylostoma ceylanicum hookworm infections in humans, Cambodia, 2012","volume":"20","author":"Inpankaew","year":"2014"},{"key":"b5","series-title":"Adv Parasitol","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1016\/S0065-308X(08)60552-7","article-title":"Hookworm infection in man","volume":"17","author":"Miller","year":"1979"},{"key":"b6","series-title":"PLoS Negl Trop Dis","first-page":"e1555","article-title":"Real-time PCR demonstrates Ancylostoma duodenale is a key factor in the etiology of severe anemia and iron deficiency in Malawian pre-school children","volume":"6","author":"Jonker","year":"2012"},{"key":"b7","series-title":"Science","first-page":"502","article-title":"Arrested development in human hookworm infections: an adaptation to a seasonally unfavourable external environment","volume":"180","author":"Schad","year":"1973"},{"key":"b8","series-title":"Am J Trop Med Hyg","first-page":"1152","article-title":"Population biology of hookworms in children in rural West Bengal. 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