{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,7]],"date-time":"2026-06-07T12:40:34Z","timestamp":1780836034071,"version":"3.54.1"},"update-to":[{"DOI":"10.1371\/journal.pntd.0013007","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2025,4,21]],"date-time":"2025-04-21T00:00:00Z","timestamp":1745193600000}}],"reference-count":51,"publisher":"Public Library of Science (PLoS)","issue":"4","license":[{"start":{"date-parts":[[2025,4,8]],"date-time":"2025-04-08T00:00:00Z","timestamp":1744070400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Shanghai Municipal Science and Technology Commission Special Foundation","award":["24DZ2203100"],"award-info":[{"award-number":["24DZ2203100"]}]},{"DOI":"10.13039\/501100013076","name":"National Major Science and Technology Projects of China","doi-asserted-by":"publisher","award":["2018ZX10734404"],"award-info":[{"award-number":["2018ZX10734404"]}],"id":[{"id":"10.13039\/501100013076","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100013076","name":"National Major Science and Technology Projects of China","doi-asserted-by":"publisher","award":["2018ZX10101002-003"],"award-info":[{"award-number":["2018ZX10101002-003"]}],"id":[{"id":"10.13039\/501100013076","id-type":"DOI","asserted-by":"publisher"}]},{"name":"the Science and Technology Leading Talent Team in Inner Mongolia Autonomous Region","award":["2022LJRC0009"],"award-info":[{"award-number":["2022LJRC0009"]}]}],"content-domain":{"domain":["www.plosntds.org"],"crossmark-restriction":false},"short-container-title":["PLoS Negl Trop Dis"],"abstract":"<jats:sec>\n<jats:title>Background<\/jats:title>\n<jats:p><jats:italic>Necator americanus<\/jats:italic> is the predominant species causing hookworm infections in humans. Despite advancements in prevention strategies, mild cases of infection still occur, highlighting the need for improved detection technology. Recombinase Polymerase Amplification (RPA) is an isothermal molecular diagnostic known for its sensitivity, speed, portability, and widespread application in detecting various pathogens. Although several molecular assays are available for <jats:italic>N. americanus<\/jats:italic>, they have limitations in detecting mild <jats:italic>N. americanus<\/jats:italic> infections.<\/jats:p>\n<\/jats:sec>\n<jats:sec>\n<jats:title>Methods<\/jats:title>\n<jats:p>Fluorescent RPA primers and probes targeting the <jats:italic>N. americanus<\/jats:italic> internal transcribed spacer 2 (<jats:italic>ITS2)<\/jats:italic> gene were developed. The method\u2019s detection limit was assessed via serial dilution of genomic DNA. Specificity was confirmed against <jats:italic>Clonorchis sinensis, Schistosoma japonicum, Fasciola hepatica, Ascaris lumbricoides, Enterobius vermicularis<\/jats:italic> and <jats:italic>Ancylostoma duodenale<\/jats:italic>. Thirty samples identified as positive by Kato-Katz, along with 11 samples identified as negative by the method, were tested to evaluate the sensitivity and specificity of fluorescent RPA. Additionally, 287 field samples were tested for validation with these methods. All positive samples were identified as either <jats:italic>N. americanus<\/jats:italic> or <jats:italic>A. duodenale<\/jats:italic>.<\/jats:p>\n<\/jats:sec>\n<jats:sec>\n<jats:title>Results<\/jats:title>\n<jats:p>This study successfully developed a fluorescent RPA assay targeting the <jats:italic>ITS2<\/jats:italic> gene of <jats:italic>N. americanus<\/jats:italic>. The length of the amplified fragment was 237\u2009bp. Optimized conditions were achieved, resulting in a minimum detection limit of 1fg\/\u00b5L, with no cross-reactivity with other pathogens. In laboratory validation, the fluorescent RPA assay demonstrated 100% sensitivity (30\/30) and 100% specificity (11\/11) compared to the Kato-Katz, and 100% sensitivity (29\/29) and 91.7% specificity (11\/12) when compared to the semi-nested PCR. In field validation using human fecal samples, the fluorescent RPA assay showed a sensitivity of 90.0% (36\/40) and a specificity of 91.1% (225\/247) compared to the Kato-Katz. And the sensitivity of the fluorescent RPA method compared to the semi-nested PCR method was 100% (34\/34), while the specificity was 90.5% (229\/252).<\/jats:p>\n<\/jats:sec>\n<jats:sec>\n<jats:title>Conclusions<\/jats:title>\n<jats:p>The fluorescent RPA assay presents a rapid and dependable method for detecting <jats:italic>N. americanus<\/jats:italic> in fecal samples. Its high sensitivity and specificity provide significant utility for field surveillance and early identification of <jats:italic>N. americanus<\/jats:italic> infections. This advancement could facilitate the rapid molecular diagnosis of <jats:italic>N. americanus<\/jats:italic> disease in hookworm-endemic regions.<\/jats:p>\n<\/jats:sec>","DOI":"10.1371\/journal.pntd.0013007","type":"journal-article","created":{"date-parts":[[2025,4,8]],"date-time":"2025-04-08T18:25:14Z","timestamp":1744136714000},"page":"e0013007","update-policy":"https:\/\/doi.org\/10.1371\/journal.pntd.corrections_policy","source":"Crossref","is-referenced-by-count":2,"title":["Development and evaluation of fluorescent recombinase polymerase amplification (RPA)-based method for rapid detection of Necator americanus"],"prefix":"10.1371","volume":"19","author":[{"given":"Jia-Rui","family":"Liang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shu-Ning","family":"Yan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Han-Yin","family":"Yang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuo","family":"Yang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu-Juan","family":"Shen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Le-Le","family":"Huo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu-Chun","family":"Cai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zi-Ran","family":"Mo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bin","family":"Zheng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-9990-2155","authenticated-orcid":true,"given":"Bin","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Hu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2025,4,8]]},"reference":[{"issue":"1","key":"pntd.0013007.ref001","doi-asserted-by":"crossref","first-page":"e72","DOI":"10.1016\/S2666-5247(21)00181-6","article-title":"Global distribution of human hookworm species and differences in their morbidity effects: a systematic review","volume":"3","author":"ACA Clements","year":"2022","journal-title":"Lancet Microbe"},{"key":"pntd.0013007.ref002","doi-asserted-by":"crossref","first-page":"16088","DOI":"10.1038\/nrdp.2016.88","article-title":"Hookworm infection","volume":"2","author":"A Loukas","year":"2016","journal-title":"Nat Rev Dis Primers"},{"issue":"9521","key":"pntd.0013007.ref003","doi-asserted-by":"crossref","first-page":"1521","DOI":"10.1016\/S0140-6736(06)68653-4","article-title":"Soil-transmitted helminth infections: ascariasis, trichuriasis, and hookworm","volume":"367","author":"J Bethony","year":"2006","journal-title":"Lancet"},{"issue":"4","key":"pntd.0013007.ref004","first-page":"268","article-title":"Human hookworms from Argentina: differential diagnosis of Necator americanus and Ancylostoma duodenale in endemic populations from Buenos Aires and Misiones","volume":"54","author":"A Servi\u00e1n","year":"2022","journal-title":"Rev Argent Microbiol"},{"issue":"10117","key":"pntd.0013007.ref005","doi-asserted-by":"crossref","first-page":"252","DOI":"10.1016\/S0140-6736(17)31930-X","article-title":"Soil-transmitted helminth infections","volume":"391","author":"PM Jourdan","year":"2018","journal-title":"Lancet"},{"issue":"12","key":"pntd.0013007.ref006","doi-asserted-by":"crossref","first-page":"e1024","DOI":"10.1016\/S2666-5247(23)00218-5","article-title":"Protective efficacy of short-term infection with Necator americanus hookworm larvae in healthy volunteers in the Netherlands: a single-centre, placebo-controlled, randomised, controlled, phase 1 trial","volume":"4","author":"M-A Hoogerwerf","year":"2023","journal-title":"Lancet Microbe"},{"issue":"1","key":"pntd.0013007.ref007","doi-asserted-by":"crossref","first-page":"e0009032","DOI":"10.1371\/journal.pntd.0009032","article-title":"Diagnosis of soil-transmitted helminths using the Kato-Katz technique: what is the influence of stirring, storage time and storage temperature on stool sample egg counts?","volume":"15","author":"F Bosch","year":"2021","journal-title":"PLoS Negl Trop Dis"},{"issue":"3","key":"pntd.0013007.ref008","doi-asserted-by":"crossref","first-page":"e39","DOI":"10.1016\/j.clinre.2017.11.008","article-title":"Human hookworm disease: enteroscopy and video capsule endoscopy features","volume":"42","author":"T Coton","year":"2018","journal-title":"Clin Res Hepatol Gastroenterol"},{"issue":"6","key":"pntd.0013007.ref009","doi-asserted-by":"crossref","first-page":"715","DOI":"10.1017\/S0031182020000220","article-title":"Use of polyclonal IgY antibodies to detect serum immune complexes in patients with active hookworm infection","volume":"147","author":"DCD Souza","year":"2020","journal-title":"Parasitology"},{"issue":"1","key":"pntd.0013007.ref010","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1007\/s00436-004-1256-8","article-title":"Infective larvae of the human hookworms Necator americanus and Ancylostoma duodenale differ in their orientation behaviour when crawling on surfaces","volume":"95","author":"W Haas","year":"2005","journal-title":"Parasitol Res"},{"issue":"2","key":"pntd.0013007.ref011","doi-asserted-by":"crossref","first-page":"e1522","DOI":"10.1371\/journal.pntd.0001522","article-title":"Epidemiological and genetic data supporting the transmission of Ancylostoma ceylanicum among human and domestic animals","volume":"6","author":"R Ngui","year":"2012","journal-title":"PLoS Negl Trop Dis"},{"issue":"3","key":"pntd.0013007.ref012","doi-asserted-by":"crossref","first-page":"e1555","DOI":"10.1371\/journal.pntd.0001555","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":"FAM Jonker","year":"2012","journal-title":"PLoS Negl Trop Dis"},{"key":"pntd.0013007.ref013","doi-asserted-by":"crossref","first-page":"380","DOI":"10.1186\/s13071-015-0994-z","article-title":"Identification of human intestinal parasites affecting an asymptomatic peri-urban Argentinian population using multi-parallel quantitative real-time polymerase chain reaction","volume":"8","author":"RO Cimino","year":"2015","journal-title":"Parasit Vectors"},{"key":"pntd.0013007.ref014","doi-asserted-by":"crossref","first-page":"574","DOI":"10.1186\/s13071-015-1183-9","article-title":"Development and evaluation of a loop mediated isothermal amplification (LAMP) technique for the detection of hookworm (Necator americanus) infection in fecal samples","volume":"8","author":"RM Mugambi","year":"2015","journal-title":"Parasit Vectors"},{"issue":"6","key":"pntd.0013007.ref015","doi-asserted-by":"crossref","first-page":"101649","DOI":"10.1016\/j.bjid.2021.101649","article-title":"Scope and limitations of a multiplex conventional PCR for the diagnosis of S. stercoralis and hookworms","volume":"25","author":"PE Fleitas","year":"2021","journal-title":"Braz J Infect Dis"},{"issue":"3","key":"pntd.0013007.ref016","doi-asserted-by":"crossref","first-page":"e2730","DOI":"10.1371\/journal.pntd.0002730","article-title":"Rapid molecular assays for the detection of yellow fever virus in low-resource settings","volume":"8","author":"C Escadafal","year":"2014","journal-title":"PLoS Negl Trop Dis"},{"key":"pntd.0013007.ref017","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1186\/1475-2875-13-99","article-title":"Rapid detection of Plasmodium falciparum with isothermal recombinase polymerase amplification and lateral flow analysis","volume":"13","author":"S Kersting","year":"2014","journal-title":"Malar J"},{"issue":"5","key":"pntd.0013007.ref018","doi-asserted-by":"crossref","first-page":"4953","DOI":"10.3390\/ijerph110504953","article-title":"Development of a recombinase polymerase amplification assay for the detection of pathogenic Leptospira","volume":"11","author":"A Ahmed","year":"2014","journal-title":"Int J Environ Res Public Health"},{"issue":"1","key":"pntd.0013007.ref019","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1016\/j.jmoldx.2013.08.003","article-title":"Sensitive and rapid detection of Chlamydia trachomatis by recombinase polymerase amplification directly from urine samples","volume":"16","author":"K Kr\u00f5lov","year":"2014","journal-title":"J Mol Diagn"},{"issue":"3","key":"pntd.0013007.ref020","doi-asserted-by":"crossref","first-page":"319","DOI":"10.3390\/pathogens11030319","article-title":"Laboratory evaluation of a basic recombinase polymerase amplification (RPA) assay for early detection of Schistosoma japonicum","volume":"11","author":"W Deng","year":"2022","journal-title":"Pathogens"},{"issue":"7","key":"pntd.0013007.ref021","doi-asserted-by":"crossref","first-page":"725","DOI":"10.1080\/14737159.2022.2109964","article-title":"Critical insight into recombinase polymerase amplification technology","volume":"22","author":"MA Munawar","year":"2022","journal-title":"Expert Rev Mol Diagn"},{"issue":"1","key":"pntd.0013007.ref022","doi-asserted-by":"crossref","first-page":"256","DOI":"10.1186\/s12917-023-03703-3","article-title":"The establishment of a recombinase polymerase amplification technique for the detection of mouse poxvirus","volume":"19","author":"Y Lian","year":"2023","journal-title":"BMC Vet Res"},{"key":"pntd.0013007.ref023","doi-asserted-by":"crossref","first-page":"105511","DOI":"10.1016\/j.actatropica.2020.105511","article-title":"Detection of Plasmodium knowlesi using recombinase polymerase amplification (RPA) combined with SYBR Green I","volume":"208","author":"MY Lai","year":"2020","journal-title":"Acta Trop"},{"issue":"1","key":"pntd.0013007.ref024","doi-asserted-by":"crossref","first-page":"514","DOI":"10.1186\/s13071-019-3755-6","article-title":"Development of a recombinase polymerase amplification (RPA) fluorescence assay for the detection of Schistosoma haematobium","volume":"12","author":"P Rostron","year":"2019","journal-title":"Parasit Vectors"},{"issue":"9","key":"pntd.0013007.ref025","doi-asserted-by":"crossref","first-page":"e0009782","DOI":"10.1371\/journal.pntd.0009782","article-title":"Sensitive and semiquantitative detection of soil-transmitted helminth infection in stool using a recombinase polymerase amplification-based assay","volume":"15","author":"JL Cantera","year":"2021","journal-title":"PLoS Negl Trop Dis"},{"key":"pntd.0013007.ref026","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/j.trac.2017.10.015","article-title":"Recombinase polymerase amplification: basics, applications and recent advances","volume":"98","author":"IM Lobato","year":"2018","journal-title":"Trends Analyt Chem"},{"issue":"3","key":"pntd.0013007.ref027","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1006\/mcpr.1996.0094","article-title":"Differentiation of Oesophagostomum bifurcum from Necator americanus by PCR using genetic markers in spacer ribosomal DNA","volume":"11","author":"A Romstad","year":"1997","journal-title":"Mol Cell Probes"},{"issue":"9","key":"pntd.0013007.ref028","doi-asserted-by":"crossref","first-page":"726","DOI":"10.1046\/j.1365-3156.2001.00770.x","article-title":"Determining the prevalence of Oesophagostomum bifurcum and Necator americanus infections using specific PCR amplification of DNA from faecal samples","volume":"6","author":"JJ Verweij","year":"2001","journal-title":"Trop Med Int Health"},{"issue":"6","key":"pntd.0013007.ref029","doi-asserted-by":"crossref","first-page":"574","DOI":"10.1111\/j.1365-3156.2005.01440.x","article-title":"Polymerase chain reaction-based differential diagnosis of Ancylostoma duodenale and Necator americanus infections in humans in northern Ghana","volume":"10","author":"JM de Gruijter","year":"2005","journal-title":"Trop Med Int Health"},{"issue":"9","key":"pntd.0013007.ref030","doi-asserted-by":"crossref","first-page":"e0004922","DOI":"10.1371\/journal.pntd.0004922","article-title":"The global economic and health burden of human hookworm infection","volume":"10","author":"SM Bartsch","year":"2016","journal-title":"PLoS Negl Trop Dis"},{"issue":"2","key":"pntd.0013007.ref031","doi-asserted-by":"crossref","first-page":"e0007155","DOI":"10.1371\/journal.pntd.0007155","article-title":"Rapid detection of Mycobacterium ulcerans with isothermal recombinase polymerase amplification assay","volume":"13","author":"M Frimpong","year":"2019","journal-title":"PLoS Negl Trop Dis"},{"key":"pntd.0013007.ref032","doi-asserted-by":"crossref","first-page":"1004","DOI":"10.3389\/fmicb.2019.01004","article-title":"Rapid and sensitive recombinase polymerase amplification combined with lateral flow strip for detecting African Swine fever virus","volume":"10","author":"F Miao","year":"2019","journal-title":"Front Microbiol"},{"issue":"22","key":"pntd.0013007.ref033","doi-asserted-by":"crossref","first-page":"19953","DOI":"10.1021\/acsomega.9b02886","article-title":"Applying a linear amplification strategy to recombinase polymerase amplification for uniform DNA library amplification","volume":"4","author":"J Lee","year":"2019","journal-title":"ACS Omega"},{"issue":"13","key":"pntd.0013007.ref034","doi-asserted-by":"crossref","first-page":"1221","DOI":"10.1017\/S0031182023000975","article-title":"Development and application of recombinase polymerase amplification assay for rapid detection of Blastocystis sp","volume":"150","author":"X Mei","year":"2023","journal-title":"Parasitology"},{"key":"pntd.0013007.ref035","doi-asserted-by":"crossref","first-page":"100046","DOI":"10.1016\/j.crpvbd.2021.100046","article-title":"Comparative studies on faecal egg counting techniques used for the detection of gastrointestinal parasites of equines: a systematic review","volume":"1","author":"A Ghafar","year":"2021","journal-title":"Curr Res Parasitol Vector Borne Dis"},{"issue":"1","key":"pntd.0013007.ref036","doi-asserted-by":"crossref","first-page":"e0009032","DOI":"10.1371\/journal.pntd.0009032","article-title":"Diagnosis of soil-transmitted helminths using the Kato-Katz technique: what is the influence of stirring, storage time and storage temperature on stool sample egg counts?","volume":"15","author":"F Bosch","year":"2021","journal-title":"PLoS Negl Trop Dis"},{"key":"pntd.0013007.ref037","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/j.mimet.2014.04.007","article-title":"Nested-quantitative PCR approach with improved sensitivity for the detection of low titer levels of Candidatus Liberibacter asiaticus in the Asian citrus psyllid, Diaphorina citri Kuwayama","volume":"102","author":"MR Coy","year":"2014","journal-title":"J Microbiol Methods"},{"issue":"1","key":"pntd.0013007.ref038","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1039\/C8AN01621F","article-title":"Review: a comprehensive summary of a decade development of the recombinase polymerase amplification","volume":"144","author":"J Li","year":"2018","journal-title":"Analyst"},{"issue":"2","key":"pntd.0013007.ref039","doi-asserted-by":"crossref","first-page":"e0007155","DOI":"10.1371\/journal.pntd.0007155","article-title":"Rapid detection of Mycobacterium ulcerans with isothermal recombinase polymerase amplification assay","volume":"13","author":"M Frimpong","year":"2019","journal-title":"PLoS Negl Trop Dis"},{"issue":"6","key":"pntd.0013007.ref040","doi-asserted-by":"crossref","first-page":"850","DOI":"10.3390\/f13060850","article-title":"Development and comparison of seminested PCR, qPCR, and LAMP for the rapid detection of Arthrinium phaeospermum, the Causal Agent of Bamboo Blight","volume":"13","author":"H Zhang","year":"2022","journal-title":"Forests"},{"key":"pntd.0013007.ref041","unstructured":"Neidler S. What are the differences between PCR, RT-PCR, qPCR, and RT-qPCR[M]\/\/Enzo. 2018."},{"issue":"1","key":"pntd.0013007.ref042","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1007\/s00216-012-6399-3","article-title":"Application of quantitative PCR for the detection of microorganisms in water","volume":"405","author":"M Botes","year":"2013","journal-title":"Anal Bioanal Chem"},{"issue":"6","key":"pntd.0013007.ref043","doi-asserted-by":"crossref","first-page":"1041","DOI":"10.4269\/ajtmh.12-0726","article-title":"A novel, multi-parallel, real-time polymerase chain reaction approach for eight gastrointestinal parasites provides improved diagnostic capabilities to resource-limited at-risk populations","volume":"88","author":"R Mejia","year":"2013","journal-title":"Am J Trop Med Hyg"},{"key":"pntd.0013007.ref044","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1016\/j.actatropica.2017.06.011","article-title":"Hookworm infections among migrant workers in Malaysia: molecular identification of Necator americanus and Ancylostoma duodenale","volume":"173","author":"N Sahimin","year":"2017","journal-title":"Acta Trop"},{"issue":"5","key":"pntd.0013007.ref045","doi-asserted-by":"crossref","first-page":"e0007372","DOI":"10.1371\/journal.pntd.0007372","article-title":"Formative research to inform development of a new diagnostic for soil-transmitted helminths: going beyond the laboratory to ensure access to a needed product","volume":"13","author":"HL Storey","year":"2019","journal-title":"PLoS Negl Trop Dis"},{"issue":"1","key":"pntd.0013007.ref046","doi-asserted-by":"crossref","first-page":"e0009032","DOI":"10.1371\/journal.pntd.0009032","article-title":"Diagnosis of soil-transmitted helminths using the Kato-Katz technique: what is the influence of stirring, storage time and storage temperature on stool sample egg counts?","volume":"15","author":"F Bosch","year":"2021","journal-title":"PLoS Negl Trop Dis"},{"issue":"5","key":"pntd.0013007.ref047","first-page":"666","article-title":"Analysis of the national prevalence of soil-transmitted nematode infections in China","volume":"39","author":"H Zhu","year":"2019","journal-title":"Chin J Parasitol Parasitic Dis"},{"issue":"3","key":"pntd.0013007.ref048","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1017\/S0007485311000575","article-title":"Suction sampling as a significant source of error in molecular analysis of predator diets","volume":"102","author":"RA King","year":"2012","journal-title":"Bull Entomol Res"},{"issue":"7","key":"pntd.0013007.ref049","doi-asserted-by":"crossref","first-page":"e0003936","DOI":"10.1371\/journal.pntd.0003936","article-title":"Quantification of Leishmania (Viannia) Kinetoplast DNA in Ulcers of Cutaneous Leishmaniasis Reveals Inter-site and Inter-sampling Variability in Parasite Load","volume":"9","author":"M Su\u00e1rez","year":"2015","journal-title":"PLoS Negl Trop Dis"},{"issue":"10","key":"pntd.0013007.ref050","doi-asserted-by":"crossref","first-page":"e0005953","DOI":"10.1371\/journal.pntd.0005953","article-title":"Estimating sensitivity of the Kato-Katz technique for the diagnosis of Schistosoma mansoni and hookworm in relation to infection intensity","volume":"11","author":"O B\u00e4renbold","year":"2017","journal-title":"PLoS Negl Trop Dis"},{"issue":"3","key":"pntd.0013007.ref051","doi-asserted-by":"crossref","first-page":"535","DOI":"10.4269\/ajtmh.13-0268","article-title":"Diagnostic accuracy of Kato-Katz, FLOTAC, Baermann, and PCR methods for the detection of light-intensity hookworm and Strongyloides stercoralis infections in Tanzania","volume":"90","author":"S Knopp","year":"2014","journal-title":"Am J Trop Med Hyg"}],"updated-by":[{"DOI":"10.1371\/journal.pntd.0013007","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2025,4,21]],"date-time":"2025-04-21T00:00:00Z","timestamp":1745193600000}}],"container-title":["PLOS Neglected Tropical Diseases"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pntd.0013007","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,4,21]],"date-time":"2025-04-21T18:11:59Z","timestamp":1745259119000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pntd.0013007"}},"subtitle":[],"editor":[{"given":"Timir","family":"Tripathi","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2025,4,8]]},"references-count":51,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2025,4,8]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pntd.0013007","relation":{"new_version":[{"id-type":"doi","id":"10.1371\/journal.pntd.0013007","asserted-by":"object"}]},"ISSN":["1935-2735"],"issn-type":[{"value":"1935-2735","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,4,8]]}}}