{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,26]],"date-time":"2026-10-26T17:59:36Z","timestamp":1793037576652,"version":"4.5.0"},"reference-count":51,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,5,13]],"date-time":"2024-05-13T00:00:00Z","timestamp":1715558400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001713","name":"European and Developing Countries Clinical Trials Partnership","doi-asserted-by":"publisher","award":["RIA2017NCT-1845"],"award-info":[{"award-number":["RIA2017NCT-1845"]}],"id":[{"id":"10.13039\/501100001713","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Parasitol."],"abstract":"<jats:p>Detection of <jats:italic>Schistosoma<\/jats:italic> spp. DNA in gynaecological samples by quantitative real-time polymerase chain reaction (qPCR) is considered to be the reference diagnostic test for female genital schistosomiasis (FGS). However, qPCR needs expensive laboratory procedures and highly trained technicians. Loop-mediated amplification (LAMP) is a more field-friendly isothermal procedure for the detection of parasite-specific DNA, but it still requires electrically powered equipment. Here, we validated a <jats:italic>Schistosoma haematobium<\/jats:italic>-specific Sh-LAMP procedure and tested a fully instrument-free isothermal amplification using a novel low-cost, and reusable Temperature-cup (T-cup) device. Specific primers were selected based on published assays, targeting the ribosomal intergenic spacer (IGS) region of <jats:italic>S. haematobium<\/jats:italic>. Technical validation of the IGS-Sh-LAMP was performed using 20 negative controls, including DNA extracts of soil-transmitted helminths and <jats:italic>S. mansoni<\/jats:italic>, and a 10-fold dilution series (10<jats:sup>0<\/jats:sup>\u201310<jats:sup>\u22123<\/jats:sup>) of DNA extracted from a single <jats:italic>S. haematobium<\/jats:italic> egg (n=4). For clinical validation, the IGS-Sh-LAMP was tested on 125 DNA samples extracted from vaginal swabs of a previous FGS study in Madagascar. Results were compared with the quantification cycle value (Cq) of the standard ITS-2 targeting qPCR. Single <jats:italic>S. haematobium<\/jats:italic> egg DNA up to a 10<jats:sup>\u20132<\/jats:sup> dilution and an ITS-2 Cq &amp;lt;35 tested positive in the IGS-Sh-LAMP. The specificity was found to be excellent (100%). In the clinical samples, IGS-Sh-LAMP showed comparable results with the qPCR, with 35.2% and 33.6% positives, respectively, and a concordance of 79.2% (99\/125). Of the 12 false-negatives, 5 corresponded to the 7 qPCR positive samples with very low DNA levels (Cq \u226535). On the other hand, IGS-Sh-LAMP detected 14 additional cases that were not detected by qPCR. The T-cup IGS-Sh-LAMP performance was evaluated in a representative sub-selection (n=10) of IGS-Sh-LAMP positive clinical samples. The T-cup IGS-Sh-LAMP was found to be a very user-friendly method, but in different runs, it missed 1 to 4 of the 10 IGS-Sh-LAMP positive samples, specifically those with a low DNA load. Our results show that the IGS-Sh-LAMP is a suitable alternative to the ITS-2 qPCR for the diagnosis of FGS in gynaecological samples, with high potential for the T-cup as a fully instrument-free isothermal amplification device for point-of-care diagnosis in low-resource settings.<\/jats:p>","DOI":"10.3389\/fpara.2024.1297310","type":"journal-article","created":{"date-parts":[[2024,5,13]],"date-time":"2024-05-13T09:55:42Z","timestamp":1715594142000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":13,"title":["Next step towards point-of-care molecular diagnosis of female genital schistosomiasis (FGS): evaluation of an instrument-free LAMP procedure"],"prefix":"10.3389","volume":"3","author":[{"given":"Kim J. M.","family":"van Bergen","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eric A.T.","family":"Brienen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bodo S.","family":"Randrianasolo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Charles E.","family":"Ramarokoto","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Leutscher","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eyrun F.","family":"Kjetland","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Angela","family":"van Diepen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Floris","family":"Dekker","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vittorio","family":"Saggiomo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aldrik H.","family":"Velders","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lisette","family":"van Lieshout","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,5,13]]},"reference":[{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0010276","article-title":"Validation of the isothermal Schistosoma haematobium Recombinase Polymerase Amplification (RPA) assay, coupled with simplified sample preparation, for diagnosing female genital schistosomiasis using cervicovaginal lavage and vaginal self-swab samples","volume":"16","author":"Archer","year":"2022","journal-title":"PloS Negl. Trop. Dis."},{"key":"B51","doi-asserted-by":"publisher","first-page":"819","DOI":"10.1017\/S0031182021000342","article-title":"Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati","volume":"148","author":"Avila","year":"2021","journal-title":"Parasitology"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0001961","article-title":"Break Out: urogenital schistosomiasis and Schistosoma haematobium infection in the post-genomic era","volume":"7","author":"Brindley","year":"2013","journal-title":"PloS Negl. Trop. Dis."},{"key":"B9","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/bs.apar.2021.12.003","article-title":"An update on female and male genital schistosomiasis and a call to integrate efforts to escalate diagnosis, treatment and awareness in endemic and non-endemic settings: The time is now","volume":"115","author":"Bustinduy","year":"2022","journal-title":"Adv. Parasitol."},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0002413","article-title":"Schistosoma haematobium-specific real-time PCR for diagnosis of urogenital schistosomiasis in serum samples of international travelers and migrants","volume":"7","author":"Cnops","year":"2013","journal-title":"PloS Negl. Trop. Dis."},{"key":"B40","doi-asserted-by":"publisher","first-page":"1208","DOI":"10.1111\/j.1365-3156.2012.03060.x","article-title":"Schistosoma real-time PCR as diagnostic tool for international travellers and migrants","volume":"17","author":"Cnops","year":"2012","journal-title":"Trop. Med. Int. Health"},{"key":"B2","doi-asserted-by":"publisher","first-page":"2253","DOI":"10.1016\/S0140-6736(13)61949-2","article-title":"Human schistosomiasis","volume":"383","author":"Colley","year":"2014","journal-title":"Lancet (London England)"},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0008231","article-title":"First international external quality assessment scheme of nucleic acid amplification tests for the detection of Schistosoma and soil-transmitted helminths, including Strongyloides: A pilot study","volume":"14","author":"Cools","year":"2020","journal-title":"PloS Negl. Trop. Dis."},{"key":"B33","doi-asserted-by":"publisher","first-page":"1308","DOI":"10.3390\/jcm10061308","article-title":"Application of a Genus-Specific LAMP Assay for Schistosome Species to Detect Schistosoma haematobium x Schistosoma bovis Hybrids","volume":"10","author":"Crego-Vicente","year":"2021","journal-title":"J. Clin. Med."},{"key":"B6","doi-asserted-by":"publisher","first-page":"615","DOI":"10.2471\/BLT.20.252270","article-title":"Integration of prevention and control measures for female genital schistosomiasis, HIV and cervical cancer","volume":"98","author":"Engels","year":"2020","journal-title":"Bull. World Health Organ"},{"key":"B44","doi-asserted-by":"publisher","first-page":"41","DOI":"10.1111\/j.1574-695X.2011.00785.x","article-title":"Robustness of a loop-mediated isothermal amplification reaction for diagnostic applications","volume":"62","author":"Francois","year":"2011","journal-title":"FEMS Immunol. Med. Microbiol."},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0003963","article-title":"The rapid-heat LAMPellet method: A potential diagnostic method for human urogenital schistosomiasis","volume":"9","author":"Gandasegui","year":"2015","journal-title":"PloS Negl. Trop. Dis."},{"key":"B50","doi-asserted-by":"publisher","first-page":"14744","DOI":"10.1038\/s41598-019-51342-2","article-title":"Progress in loop-mediated isothermal amplification assay for detection of Schistosoma mansoni DNA: towards a ready-to-use test","volume":"9","author":"Garc\u00eda-Bernalt Diego","year":"2019","journal-title":"Sci. Rep."},{"key":"B28","doi-asserted-by":"publisher","first-page":"511","DOI":"10.3390\/jcm10030511","article-title":"Loop-mediated isothermal amplification in schistosomiasis","volume":"10","author":"Garc\u00eda-Bernalt Diego","year":"2021","journal-title":"J. Clin. Med."},{"key":"B29","doi-asserted-by":"publisher","first-page":"438","DOI":"10.3390\/diagnostics11030521","article-title":"LAMP in neglected tropical diseases: A focus on parasites","volume":"11","author":"Garc\u00eda-Bernalt Diego","year":"2021","journal-title":"Diagnostics (Basel)"},{"key":"B25","doi-asserted-by":"publisher","first-page":"344","DOI":"10.4269\/ajtmh.2012.12-0208","article-title":"Evaluation of loop-mediated isothermal amplification suitable for molecular monitoring of schistosome-infected snails in field laboratories","volume":"88","author":"Hamburger","year":"2013","journal-title":"Am. J. Trop. Med. Hyg"},{"key":"B39","doi-asserted-by":"publisher","first-page":"907","DOI":"10.4269\/ajtmh.2001.65.907","article-title":"Polymerase chain reaction assay based on a highly repeated sequence of Schistosoma haematobium: a potential tool for monitoring schistosome-infested water","volume":"65","author":"Hamburger","year":"2001","journal-title":"Am. J. Trop. Med. Hyg"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0000412","article-title":"Neglected tropical diseases in sub-saharan Africa: review of their prevalence, distribution, and disease burden","volume":"3","author":"Hotez","year":"2009","journal-title":"PloS Negl. Trop. Dis."},{"key":"B21","doi-asserted-by":"publisher","first-page":"2616","DOI":"10.1128\/JCM.41.6.2616-2622.2003","article-title":"Loop-mediated isothermal amplification for direct detection of Mycobacterium tuberculosis complex, M. avium, and M. intracellulare in sputum samples","volume":"41","author":"Iwamoto","year":"2003","journal-title":"J. Clin. Microbiol."},{"key":"B14","doi-asserted-by":"publisher","first-page":"1050","DOI":"10.4269\/ajtmh.2009.09-0081","article-title":"Schistosomiasis PCR in vaginal lavage as an indicator of genital Schistosoma haematobium infection in rural Zimbabwean women","volume":"81","author":"Kjetland","year":"2009","journal-title":"Am. J. Trop. Med. Hyg"},{"key":"B12","doi-asserted-by":"publisher","first-page":"58","DOI":"10.1016\/j.pt.2011.10.008","article-title":"A review of female genital schistosomiasis","volume":"28","author":"Kjetland","year":"2012","journal-title":"Trends Parasitol."},{"key":"B42","doi-asserted-by":"publisher","first-page":"166","DOI":"10.1186\/s12936-021-03707-0","article-title":"Validation of SYBR green I based closed-tube loop-mediated isothermal amplification (LAMP) assay for diagnosis of knowlesi malaria","volume":"20","author":"Lai","year":"2021","journal-title":"Malar J."},{"key":"B7","doi-asserted-by":"publisher","first-page":"775","DOI":"10.1086\/591127","article-title":"Coexistence of urogenital schistosomiasis and sexually transmitted infection in women and men living in an area where Schistosoma haematobium is endemic","volume":"47","author":"Leutscher","year":"2008","journal-title":"Clin. Infect. Dis."},{"key":"B11","doi-asserted-by":"publisher","first-page":"380","DOI":"10.1186\/s12913-018-3207-6","article-title":"Operational assessment of point-of-care diagnostics in rural primary healthcare clinics of KwaZulu-Natal, South Africa: a cross-sectional survey","volume":"18","author":"Mashamba-Thompson","year":"2018","journal-title":"BMC Health Serv. Res."},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2022.1043596","article-title":"Development of real-time and lateral flow recombinase polymerase amplification assays for rapid detection of Schistosoma mansoni","volume":"13","author":"Mesquita","year":"2022","journal-title":"Front. Microbiol."},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0003959","article-title":"Is PCR the next reference standard for the diagnosis of Schistosoma in stool? A comparison with microscopy in Senegal and Kenya","volume":"9","author":"Meurs","year":"2015","journal-title":"PloS Negl. Trop. Dis."},{"key":"B20","doi-asserted-by":"publisher","first-page":"43","DOI":"10.1080\/14737159.2021.1873769","article-title":"LAMP diagnostics at the point-of-care: emerging trends and perspectives for the developer community","volume":"21","author":"Moehling","year":"2021","journal-title":"Expert Rev. Mol. Diagnostics"},{"key":"B46","doi-asserted-by":"publisher","first-page":"367","DOI":"10.3390\/bios13030367","article-title":"Low-temperature loop-mediated isothermal amplification operating at physiological temperature","volume":"13","author":"Nam","year":"2023","journal-title":"Biosensors"},{"key":"B47","doi-asserted-by":"publisher","first-page":"5916","DOI":"10.1038\/s41598-024-55241-z","article-title":"Evaluation of molecular inhibitors of loop-mediated isothermal amplification (LAMP)","volume":"14","author":"Nwe","year":"2024","journal-title":"Sci. Rep."},{"key":"B36","doi-asserted-by":"publisher","first-page":"625","DOI":"10.1179\/136485908X337490","article-title":"Application of a circulating-cathodic-antigen (CCA) strip test and real-time PCR, in comparison with microscopy, for the detection of Schistosoma haematobium in urine samples from Ghana","volume":"102","author":"Obeng","year":"2008","journal-title":"Ann. Trop. Med. Parasitol."},{"key":"B43","doi-asserted-by":"publisher","first-page":"6618394","DOI":"10.1155\/2021\/6618394","article-title":"Prevalence, risk factors, and coinfection of urogenital schistosomiasis and soil-transmitted helminthiasis among primary school children in biase, southern Nigeria","volume":"2021","author":"Opara","year":"2021","journal-title":"J. Parasitol. Res."},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pmed.1001306","article-title":"Point-of-care testing for infectious diseases: diversity, complexity, and barriers in low- and middle-income countries","volume":"9","author":"Pai","year":"2012","journal-title":"PloS Med."},{"key":"B10","doi-asserted-by":"publisher","first-page":"1062","DOI":"10.1111\/j.1469-0691.2010.03279.x","article-title":"Point-of-care tests for diagnosing infections in the developing world","volume":"16","author":"Peeling","year":"2010","journal-title":"Clin. Microbiol. Infection"},{"key":"B15","doi-asserted-by":"publisher","first-page":"105363","DOI":"10.1016\/j.actatropica.2020.105363","article-title":"Detection of Schistosoma DNA in genital specimens and urine: A comparison between five female African study populations originating from S. haematobium and\/or S. mansoni endemic areas","volume":"204","author":"Pillay","year":"2020","journal-title":"Acta Tropica"},{"key":"B13","doi-asserted-by":"publisher","first-page":"233","DOI":"10.4269\/ajtmh.2001.65.233","article-title":"Diagnosis of genital cervical schistosomiasis: comparison of cytological, histopathological and parasitological examination","volume":"65","author":"Poggensee","year":"2001","journal-title":"Am. J. Trop. Med. Hyg"},{"key":"B22","doi-asserted-by":"publisher","first-page":"303","DOI":"10.1373\/clinchem.2005.057901","article-title":"Sensitive and inexpensive molecular test for falciparum malaria: detecting Plasmodium falciparum DNA directly from heat-treated blood by loop-mediated isothermal amplification","volume":"52","author":"Poon","year":"2006","journal-title":"Clin. Chem."},{"key":"B8","doi-asserted-by":"publisher","first-page":"691","DOI":"10.1186\/s12879-021-06380-5","article-title":"Association between cervical dysplasia and female genital schistosomiasis diagnosed by genital PCR in Zambian women","volume":"21","author":"Rafferty","year":"2021","journal-title":"BMC Infect. Dis."},{"key":"B35","doi-asserted-by":"publisher","first-page":"275","DOI":"10.1093\/infdis\/jiv035","article-title":"Gynecological manifestations, histopathological findings, and schistosoma-specific polymerase chain reaction results among women with Schistosoma haematobium infection: a cross-sectional study in Madagascar","volume":"212","author":"Randrianasolo","year":"2015","journal-title":"J. Infect. Dis."},{"key":"B48","doi-asserted-by":"publisher","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":"Rostron","year":"2019","journal-title":"Parasites Vectors"},{"key":"B49","doi-asserted-by":"publisher","first-page":"7043","DOI":"10.1038\/s41598-022-11144-5","article-title":"A lyophilized colorimetric RT-LAMP test kit for rapid, low-cost, at-home molecular testing of SARS-CoV-2 and other pathogens","volume":"12","author":"Song","year":"2022","journal-title":"Sci. Rep."},{"key":"B5","doi-asserted-by":"publisher","first-page":"105524","DOI":"10.1016\/j.actatropica.2020.105524","article-title":"Beyond the barrier: Female Genital Schistosomiasis as a potential risk factor for HIV-1 acquisition","volume":"209","author":"Sturt","year":"2020","journal-title":"Acta tropica"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0008337","article-title":"Genital self-sampling compared with cervicovaginal lavage for the diagnosis of female genital schistosomiasis in Zambian women: The BILHIV study","volume":"14","author":"Sturt","year":"2020","journal-title":"PloS Negl. Trop. Dis."},{"key":"B23","doi-asserted-by":"publisher","first-page":"578","DOI":"10.4269\/ajtmh.2009.09-0145","article-title":"Sensitive, specific, and rapid detection of Leishmania donovani DNA by loop-mediated isothermal amplification","volume":"81","author":"Takagi","year":"2009","journal-title":"Am. J. Trop. Med. Hyg"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0011465","article-title":"Assessing the prevalence of Female Genital Schistosomiasis and comparing the acceptability and performance of health worker-collected and self-collected cervical-vaginal swabs using PCR testing among women in North-Western Tanzania: The ShWAB study","volume":"17","author":"Ursini","year":"2023","journal-title":"PloS Negl. Trop. Dis."},{"key":"B31","doi-asserted-by":"publisher","first-page":"2100078","DOI":"10.1002\/gch2.202100078","article-title":"T-cup: A cheap, rapid, and simple home device for isothermal nucleic acid amplification","volume":"6","author":"Velders","year":"2022","journal-title":"Global Challenges"},{"key":"B30","doi-asserted-by":"publisher","first-page":"93","DOI":"10.1186\/s13104-018-3197-9","article-title":"Loop-mediated isothermal amplification (LAMP) shield for Arduino DNA detection","volume":"11","author":"Velders","year":"2018","journal-title":"BMC Res. Notes"},{"key":"B45","doi-asserted-by":"publisher","first-page":"112766","DOI":"10.1101\/2020.04.21.052530v1.full","article-title":"One-pot detection of COVID-19 with real-time reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay and visual RT-LAMP assay","volume":"172","author":"Wang","year":"2020","journal-title":"bioRxiv"},{"key":"B26","doi-asserted-by":"publisher","first-page":"164","DOI":"10.1186\/1756-3305-4-164","article-title":"Application of DNA-based diagnostics in detection of schistosomal DNA in early infection and after drug treatment","volume":"4","author":"Wang","year":"2011","journal-title":"Parasites Vectors"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0000422","article-title":"Diagnosing schistosomiasis by detection of cell-free parasite DNA in human plasma","volume":"3","author":"Wichmann","year":"2009","journal-title":"PloS Negl. Trop. Dis."},{"key":"B1","first-page":"621","volume-title":"Schistosomiasis and soiltransmitted helminthiases: progress report, 2021. Weekly Epidemiological Record","year":"2022"},{"key":"B19","doi-asserted-by":"publisher","first-page":"107","DOI":"10.1146\/annurev.bioeng.10.061807.160524","article-title":"Point-of-care diagnostics for global health","volume":"10","author":"Yager","year":"2008","journal-title":"Annu. Rev. Biomed. Eng."}],"container-title":["Frontiers in Parasitology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fpara.2024.1297310\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,5,13]],"date-time":"2024-05-13T09:55:49Z","timestamp":1715594149000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fpara.2024.1297310\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,5,13]]},"references-count":51,"alternative-id":["10.3389\/fpara.2024.1297310"],"URL":"https:\/\/doi.org\/10.3389\/fpara.2024.1297310","relation":{},"ISSN":["2813-2424"],"issn-type":[{"value":"2813-2424","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,5,13]]},"article-number":"1297310"}}