{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,4]],"date-time":"2026-06-04T22:31:13Z","timestamp":1780612273453,"version":"3.54.1"},"reference-count":44,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2018,8,20]],"date-time":"2018-08-20T00:00:00Z","timestamp":1534723200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100002873","name":"Chulalongkorn University","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002873","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002873","name":"Chulalongkorn University","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002873","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Clinical Laboratory Analysis"],"published-print":{"date-parts":[[2019,2]]},"abstract":"Abstract<\/jats:title>Background<\/jats:title>Rapid diagnosis of Mycobacterium tuberculosis<\/jats:italic> (Mtb<\/jats:italic>) is key to controlling the spread of tuberculosis, which is a global health concern. In this study, isothermal recombinase polymerase amplification (RPA<\/jats:styled-content>) was developed to detect specific targets of Mtb<\/jats:italic>, IS<\/jats:styled-content>6110 and IS<\/jats:styled-content>1081. Additionally, SYBR<\/jats:styled-content> Green I was used for endpoint detection of the RPA<\/jats:styled-content> products by the naked eye.<\/jats:p><\/jats:sec>Method<\/jats:title>A total of 146 genomic Mtb <\/jats:italic>DNA<\/jats:styled-content> samples and 24 genomic nontuberculous mycobacteria (NTM<\/jats:styled-content>) DNA<\/jats:styled-content> samples were amplified at IS<\/jats:styled-content>6110 and IS<\/jats:styled-content>1081 by RPA<\/jats:styled-content>. After a complete amplification, the RPA<\/jats:styled-content> amplicons were examined by agarose gel electrophoresis (RPA<\/jats:styled-content>\u2010AGE<\/jats:styled-content>) and SYBR<\/jats:styled-content> Green I (RPA<\/jats:styled-content>\u2010S) assays. The performance of the RPA<\/jats:styled-content> assays was evaluated by comparing them to a conventional PCR<\/jats:styled-content>.<\/jats:p><\/jats:sec>Results<\/jats:title>The RPA<\/jats:styled-content> assay demonstrated to have a good capability to differentiate Mtb<\/jats:italic> from NTM<\/jats:styled-content> with a very short turnaround time at a constant temperature. Compared to conventional PCR<\/jats:styled-content>, the sensitivities and specificities of RPA<\/jats:styled-content>\u2010AGE<\/jats:styled-content> for IS<\/jats:styled-content>6110 and IS<\/jats:styled-content>1081 were 100%. The specificity of RPA<\/jats:styled-content>\u2010S was 100% for both targets; however, its sensitivities for IS<\/jats:styled-content>6110 and IS<\/jats:styled-content>1081 were 97.95% and 99.32%, respectively. The limits of detection of IS<\/jats:styled-content>6110 RPA<\/jats:styled-content>\u2010AGE<\/jats:styled-content> and RPA<\/jats:styled-content>\u2010S were 0.05 and 0.5\u00a0ng, respectively, while the LOD<\/jats:styled-content>s of IS<\/jats:styled-content>1081 RPA<\/jats:styled-content>\u2010AGE<\/jats:styled-content> and RPA<\/jats:styled-content>\u2010S were 0.00005 and 0.05\u00a0ng, respectively. Both RPA<\/jats:styled-content> assays showed a satisfying diagnostic specificity, with no cross\u2010reaction with other bacteria.<\/jats:p><\/jats:sec>Conclusion<\/jats:title>A rapid, sensitive, naked eye RPA<\/jats:styled-content> assay can be integrated into point\u2010of\u2010care diagnosis for Mtb<\/jats:italic> detection, especially in remote areas where laboratory instrument resources are limited.<\/jats:p><\/jats:sec>","DOI":"10.1002\/jcla.22655","type":"journal-article","created":{"date-parts":[[2018,8,22]],"date-time":"2018-08-22T15:20:54Z","timestamp":1534951254000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":45,"title":["Naked eye detection of the Mycobacterium tuberculosis<\/i> complex by recombinase polymerase amplification\u2014SYBR<\/scp> green I assays"],"prefix":"10.1002","volume":"33","author":[{"given":"Nuntita","family":"Singpanomchai","sequence":"first","affiliation":[{"name":"Program of Molecular sciences in Medical Microbiology and Immunology Department of Transfusion Medicine and Clinical Microbiology Faculty of Allied Health Sciences Chulalongkorn University Bangkok Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yukihiro","family":"Akeda","sequence":"additional","affiliation":[{"name":"Division of Infection Control and Prevention Osaka University Hospital Osaka University Osaka Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kazunori","family":"Tomono","sequence":"additional","affiliation":[{"name":"Division of Infection Control and Prevention Osaka University Hospital Osaka University Osaka Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aki","family":"Tamaru","sequence":"additional","affiliation":[{"name":"Department of Bacteriology Osaka Prefectural Institute of Public Health Osaka Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pitak","family":"Santanirand","sequence":"additional","affiliation":[{"name":"Microbiology Unit Faculty of Medicine Ramathibodi Hospital Mahidol University Bangkok Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3084-0863","authenticated-orcid":false,"given":"Panan","family":"Ratthawongjirakul","sequence":"additional","affiliation":[{"name":"Research Group of Innovative Diagnosis of Antimicrobial Resistance Department of Transfusion Medicine and Clinical Microbiology Faculty of Allied Health Sciences Chulalongkorn University Bangkok Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2018,8,20]]},"reference":[{"key":"e_1_2_6_2_1","volume-title":"Global Tuberculosis Report 2017","author":"WHO","year":"2017"},{"key":"e_1_2_6_3_1","volume-title":"Global Tuberculosis Report 2016","author":"WHO","year":"2016"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.02289-08"},{"key":"e_1_2_6_5_1","volume-title":"Core Curriculum on Tuberculosis: What the Clinician Should Know","author":"CDC","year":"2013"},{"key":"e_1_2_6_6_1","unstructured":"WHO.Internaltional standards for tuberculosis care.http:\/\/www.who.int\/tb\/publications\/ISTC_3rdEd.pdf. 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