{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T03:44:15Z","timestamp":1790307855632,"version":"4.1.0"},"update-to":[{"DOI":"10.1371\/journal.pntd.0010180","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2022,2,9]],"date-time":"2022-02-09T00:00:00Z","timestamp":1644364800000}}],"reference-count":29,"publisher":"Public Library of Science (PLoS)","issue":"1","license":[{"start":{"date-parts":[[2022,1,28]],"date-time":"2022-01-28T00:00:00Z","timestamp":1643328000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["AI137804"],"award-info":[{"award-number":["AI137804"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100005624","name":"PATH","doi-asserted-by":"publisher","award":["OPP1112376"],"award-info":[{"award-number":["OPP1112376"]}],"id":[{"id":"10.13039\/100005624","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosntds.org"],"crossmark-restriction":false},"short-container-title":["PLoS Negl Trop Dis"],"abstract":"<jats:p>\n                    Enterotoxigenic\n                    <jats:italic>E<\/jats:italic>\n                    .\n                    <jats:italic>coli<\/jats:italic>\n                    (ETEC) and\n                    <jats:italic>Shigella spp<\/jats:italic>\n                    (Shigella) are complex pathogens. The diagnostic assays currently used to detect these pathogens are elaborate or complicated, which make them difficult to apply in resource poor settings where these diseases are endemic. The culture methods used to detect Shigella are not sensitive, and the methods used to detect ETEC are only available in a few research labs. To address this gap, we developed a rapid and simple diagnostic assay\u2013\"Rapid LAMP based Diagnostic Test (RLDT).\" The six minutes sample preparation method directly from the fecal samples with lyophilized reaction strips and using established Loop-mediated Isothermal Amplification (LAMP) platform, ETEC [heat labile toxin (LT) and heat stable toxins (STh, and STp) genes] and Shigella (\n                    <jats:italic>ipaH<\/jats:italic>\n                    gene) detection was made simple, rapid (&lt;50 minutes), and inexpensive. This assay is cold chain and electricity free. Moreover, RLDT requires minimal equipment. To avoid any end user\u2019s bias, a battery-operated, handheld reader was used to read the RLDT results. The results can be read as positive\/negative or as real time amplification depending on the end user\u2019s need. The performance specifications of the RLDT assay, including analytical sensitivity and specificity, were evaluated using fecal samples spiked with ETEC and Shigella strains. The limit of detection was ~10\n                    <jats:sup>5<\/jats:sup>\n                    CFU\/gm of stool for LT, STh, and STp and ~10\n                    <jats:sup>4<\/jats:sup>\n                    CFU\/gm of stool for the\n                    <jats:italic>ipaH<\/jats:italic>\n                    gene, which corresponds to about 23 CFU and 1 CFU respectively for ETEC and Shigella per 25uL reaction within 40 minutes. The RLDT assay from stool collection to result is simple, and rapid and at the same time sufficiently sensitive. RLDT has the potential to be applied in resource poor endemic settings for the rapid diagnosis of ETEC and Shigella.\n                  <\/jats:p>","DOI":"10.1371\/journal.pntd.0010180","type":"journal-article","created":{"date-parts":[[2022,1,28]],"date-time":"2022-01-28T13:36:46Z","timestamp":1643377006000},"page":"e0010180","update-policy":"https:\/\/doi.org\/10.1371\/journal.pntd.corrections_policy","source":"Crossref","is-referenced-by-count":25,"title":["Development of a simple, rapid, and sensitive diagnostic assay for enterotoxigenic E. coli and Shigella spp applicable to endemic countries"],"prefix":"10.1371","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6446-0255","authenticated-orcid":true,"given":"Subhra","family":"Chakraborty","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sean","family":"Connor","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1404-3688","authenticated-orcid":true,"given":"Mirza","family":"Velagic","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2022,1,28]]},"reference":[{"key":"pntd.0010180.ref001","doi-asserted-by":"crossref","first-page":"465","DOI":"10.1128\/CMR.18.3.465-483.2005","article-title":"Enterotoxigenic Escherichia coli in developing countries: epidemiology, microbiology, clinical features, treatment, and prevention","volume":"18","author":"F Qadri","year":"2005","journal-title":"Clin Microbiol Rev"},{"key":"pntd.0010180.ref002","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1016\/S0140-6736(13)60844-2","article-title":"Burden and aetiology of diarrhoeal disease in infants and young children in developing countries (the Global Enteric Multicenter Study, GEMS): a prospective, case-control study","volume":"382","author":"KL Kotloff","year":"2013","journal-title":"Lancet"},{"key":"pntd.0010180.ref003","doi-asserted-by":"crossref","first-page":"497","DOI":"10.1086\/338834","article-title":"Prevalence of enteric pathogens among international travelers with diarrhea acquired in Kenya (Mombasa), India (Goa), or Jamaica (Montego Bay)","volume":"185","author":"ZD Jiang","year":"2002","journal-title":"J Infect Dis"},{"key":"pntd.0010180.ref004","doi-asserted-by":"crossref","first-page":"633","DOI":"10.1128\/JCM.02572-12","article-title":"Genotypic characterization of enterotoxigenic Escherichia coli strains causing traveler\u2019s diarrhea","volume":"51","author":"FP Rivera","year":"2013","journal-title":"J Clin Microbiol"},{"key":"pntd.0010180.ref005","doi-asserted-by":"crossref","first-page":"3241","DOI":"10.1128\/JCM.39.9.3241-3246.2001","article-title":"Concomitant infection of enterotoxigenic Escherichia coli in an outbreak of cholera caused by Vibrio cholerae O1 and O139 in Ahmedabad, India","volume":"39","author":"S Chakraborty","year":"2001","journal-title":"J Clin Microbiol"},{"key":"pntd.0010180.ref006","doi-asserted-by":"crossref","first-page":"473","DOI":"10.1023\/B:EJEP.0000027353.70796.25","article-title":"Molecular epidemiology of diarrhoeagenic Escherichia coli associated with sporadic cases and outbreaks of diarrhoea between 2000 and 2001 in India","volume":"19","author":"S Kahali","year":"2004","journal-title":"Eur J Epidemiol"},{"key":"pntd.0010180.ref007","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1111\/1574-6968.12362","article-title":"Quantitative PCR and culture evaluation for enterotoxigenic Escherichia coli (ETEC) associated diarrhea in volunteers","volume":"352","author":"BR Lindsay","year":"2014","journal-title":"FEMS Microbiol Lett"},{"key":"pntd.0010180.ref008","doi-asserted-by":"crossref","first-page":"124","DOI":"10.4269\/ajtmh.13-0383","article-title":"Development and accuracy of quantitative real-time polymerase chain reaction assays for detection and quantification of enterotoxigenic Escherichia coli (ETEC) heat labile and heat stable toxin genes in travelers\u2019 diarrhea samples","volume":"90","author":"BP Youmans","year":"2014","journal-title":"Am J Trop Med Hyg"},{"key":"pntd.0010180.ref009","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1038\/nrgastro.2013.12","article-title":"Progress and pitfalls in Shigella vaccine research","volume":"10","author":"EM Barry","year":"2013","journal-title":"Nat Rev Gastroenterol Hepatol"},{"key":"pntd.0010180.ref010","doi-asserted-by":"crossref","first-page":"933","DOI":"10.1093\/cid\/ciu468","article-title":"Shigella isolates from the global enteric multicenter study inform vaccine development","volume":"59","author":"S Livio","year":"2014","journal-title":"Clin Infect Dis"},{"key":"pntd.0010180.ref011","doi-asserted-by":"crossref","first-page":"1740","DOI":"10.1128\/JCM.02713-12","article-title":"Quantitative PCR for detection of Shigella improves ascertainment of Shigella burden in children with moderate-to-severe diarrhea in low-income countries","volume":"51","author":"B Lindsay","year":"2013","journal-title":"J Clin Microbiol"},{"key":"pntd.0010180.ref012","doi-asserted-by":"crossref","first-page":"3263","DOI":"10.1128\/JCM.01342-13","article-title":"Survey of culture, goldengate assay, universal biosensor assay, and 16S rRNA Gene sequencing as alternative methods of bacterial pathogen detection","volume":"51","author":"B Lindsay","year":"2013","journal-title":"J Clin Microbiol"},{"key":"pntd.0010180.ref013","doi-asserted-by":"crossref","first-page":"1291","DOI":"10.1016\/S0140-6736(16)31529-X","article-title":"Use of quantitative molecular diagnostic methods to identify causes of diarrhoea in children: a reanalysis of the GEMS case-control study","volume":"388","author":"J Liu","year":"2016","journal-title":"Lancet"},{"key":"pntd.0010180.ref014","doi-asserted-by":"crossref","first-page":"4784","DOI":"10.1016\/j.vaccine.2019.01.031","article-title":"Capturing the true burden of Shigella and ETEC: The way forward","volume":"37","author":"I Khalil","year":"2019","journal-title":"Vaccine"},{"key":"pntd.0010180.ref015","doi-asserted-by":"crossref","first-page":"4266","DOI":"10.1016\/j.vaccine.2021.04.018","article-title":"Enterotoxigenic Escherichia coli (ETEC) vaccines: Priority activities to enable product development, licensure, and global access","volume":"39","author":"I Khalil","year":"2021","journal-title":"Vaccine"},{"key":"pntd.0010180.ref016","doi-asserted-by":"crossref","first-page":"E63","DOI":"10.1093\/nar\/28.12.e63","article-title":"Loop-mediated isothermal amplification of DNA","volume":"28","author":"T. Notomi","year":"2000","journal-title":"Nucleic Acids Res"},{"key":"pntd.0010180.ref017","doi-asserted-by":"crossref","first-page":"414","DOI":"10.1016\/j.mimet.2006.09.024","article-title":"Rapid and sensitive detection of heat-labile I and heat-stable I enterotoxin genes of enterotoxigenic Escherichia coli by Loop-Mediated Isothermal Amplification","volume":"68","author":"A Yano","year":"2007","journal-title":"J Microbiol Methods"},{"key":"pntd.0010180.ref018","doi-asserted-by":"crossref","first-page":"1719","DOI":"10.1128\/CVI.05194-11","article-title":"Refinement of a human challenge model for evaluation of enterotoxigenic Escherichia coli vaccines","volume":"18","author":"C Harro","year":"2011","journal-title":"Clin Vaccine Immunol"},{"key":"pntd.0010180.ref019","doi-asserted-by":"crossref","first-page":"e0006442","DOI":"10.1371\/journal.pntd.0006442","article-title":"Impact of lower challenge doses of enterotoxigenic Escherichia coli on clinical outcome, intestinal colonization and immune responses in adult volunteers","volume":"12","author":"S Chakraborty","year":"2018","journal-title":"PLoS Negl Trop Dis"},{"key":"pntd.0010180.ref020","doi-asserted-by":"crossref","first-page":"103310","DOI":"10.1016\/j.ebiom.2021.103310","article-title":"Human challenge study with a Shigella bioconjugate vaccine: Analyses of clinical efficacy and correlate of protection","volume":"66","author":"KR Talaat","year":"2021","journal-title":"EBioMedicine"},{"key":"pntd.0010180.ref021","doi-asserted-by":"crossref","first-page":"472","DOI":"10.1128\/JCM.02658-12","article-title":"A Laboratory-Developed TaqMan Array Card for Simultaneous Detection of 19 Enteropathogens","volume":"51","author":"J Liu","year":"2013","journal-title":"J Clin Microbiol"},{"key":"pntd.0010180.ref022","doi-asserted-by":"crossref","first-page":"e1309","DOI":"10.1016\/S2214-109X(18)30349-8","article-title":"MAL-ED Network Investigators. Use of quantitative molecular diagnostic methods to assess the aetiology, burden, and clinical characteristics of diarrhoea in children in low-resource settings: a reanalysis of the MAL-ED cohort study","volume":"6","author":"JA Platts-Mills","year":"2018","journal-title":"Lancet Glob Health"},{"key":"pntd.0010180.ref023","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/j.mimet.2019.04.010","article-title":"Development of isothermal amplification methods for rapid and sensitive detection of heat-labile enterotoxin producing Escherichia coli","volume":"161","author":"W Liu","year":"2019","journal-title":"J Microbiol Methods"},{"key":"pntd.0010180.ref024","doi-asserted-by":"crossref","first-page":"316","DOI":"10.5582\/bst.2014.01118","article-title":"Detection methods for milk pathogenic bacteria by loop-mediated isothermal amplification","volume":"8","author":"W Yang","year":"2014","journal-title":"Biosci Trends"},{"key":"pntd.0010180.ref025","first-page":"1400","article-title":"by Multiple Endonuclease Restriction Real-Time Loop-Mediated Isothermal Amplification Technique","volume":"6","author":"Y Wang","year":"2015","journal-title":"Rapid and Sensitive Detection of Shigella spp. and Salmonella spp.Front Microbiol"},{"key":"pntd.0010180.ref026","first-page":"709","article-title":"A real-time loop-mediated isothermal amplification assay for rapid detection of Shigella species","volume":"31","author":"PS Liew","year":"2014","journal-title":"Trop Biomed"},{"key":"pntd.0010180.ref027","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1016\/j.diagmicrobio.2013.09.009","article-title":"Evaluation of colorimetric detection methods for Shigella, Salmonella, and Vibrio cholerae by loop-mediated isothermal amplification","volume":"77","author":"KW Soli","year":"2013","journal-title":"Diagn Microbiol Infect Dis"},{"key":"pntd.0010180.ref028","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1016\/j.ijfoodmicro.2011.05.004","article-title":"Development of multiplex loop-mediated isothermal amplification-RFLP (mLAMP-RFLP) to detect Salmonella spp. and Shigella spp. in milk","volume":"148","author":"Y Shao","year":"2011","journal-title":"Int J Food Microbiol"},{"key":"pntd.0010180.ref029","doi-asserted-by":"crossref","first-page":"94","DOI":"10.3389\/fmicb.2018.00094","article-title":"Detection of Shigella in Milk and Clinical Samples by Magnetic Immunocaptured-Loop-Mediated Isothermal Amplification Assay","volume":"9","author":"L Zhang","year":"2018","journal-title":"Front Microbiol"}],"updated-by":[{"DOI":"10.1371\/journal.pntd.0010180","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2022,2,9]],"date-time":"2022-02-09T00:00:00Z","timestamp":1644364800000}}],"container-title":["PLOS Neglected Tropical Diseases"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pntd.0010180","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,2,9]],"date-time":"2022-02-09T13:42:27Z","timestamp":1644414147000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pntd.0010180"}},"subtitle":[],"editor":[{"given":"Kirkby D.","family":"Tickell","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2022,1,28]]},"references-count":29,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2022,1,28]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pntd.0010180","relation":{},"ISSN":["1935-2735"],"issn-type":[{"value":"1935-2735","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,1,28]]}}}