{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,12]],"date-time":"2026-05-12T19:52:18Z","timestamp":1778615538057,"version":"3.51.4"},"reference-count":28,"publisher":"Microbiology Society","issue":"12","funder":[{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"crossref","award":["JP18K08446"],"award-info":[{"award-number":["JP18K08446"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"crossref","award":["JP19fk0108091"],"award-info":[{"award-number":["JP19fk0108091"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2020,12,1]]},"abstract":"<jats:p>\n                  <jats:bold>Introduction.<\/jats:bold> Macrolides could be a potential alternative treatment for <jats:italic>\n                     <jats:named-content content-type=\"species\">\n                        <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" ext-link-type=\"uri\" xlink:href=\"http:\/\/doi.org\/10.1601\/nm.7858\" xlink:type=\"simple\">Treponema pallidum<\/jats:ext-link>\n                     <\/jats:named-content>\n                  <\/jats:italic> infections in patients; however, macrolide-resistant <jats:italic>\n                     <jats:named-content content-type=\"species\">\n                        <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" ext-link-type=\"uri\" xlink:href=\"http:\/\/doi.org\/10.1601\/nm.7858\" xlink:type=\"simple\">T. pallidum<\/jats:ext-link>\n                     <\/jats:named-content>\n                  <\/jats:italic> is spreading rapidly worldwide.<\/jats:p>\n               <jats:p>\n                  <jats:bold>Hypothesis\/Gap Statement<\/jats:bold>. There are presently no alternatives to serological tests for syphilis that can be used to evaluate therapeutic effects due to the fact that <jats:italic>\n                     <jats:named-content content-type=\"species\">\n                        <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" ext-link-type=\"uri\" xlink:href=\"http:\/\/doi.org\/10.1601\/nm.7858\" xlink:type=\"simple\">T. pallidum<\/jats:ext-link>\n                     <\/jats:named-content>\n                  <\/jats:italic> cannot be cultured <jats:italic>in vitro<\/jats:italic>.<\/jats:p>\n               <jats:p>\n                  <jats:bold>Aim.<\/jats:bold> In this study, we constructed a method for rapidly identifying <jats:italic>\n                     <jats:named-content content-type=\"species\">\n                        <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" ext-link-type=\"uri\" xlink:href=\"http:\/\/doi.org\/10.1601\/nm.7858\" xlink:type=\"simple\">T. pallidum<\/jats:ext-link>\n                     <\/jats:named-content>\n                  <\/jats:italic> and confirming macrolide resistance by using loop-mediated isothermal amplification (LAMP) with peptide nucleic acids (PNAs).<\/jats:p>\n               <jats:p>\n                  <jats:bold>Methodology.<\/jats:bold> A set of LAMP primers was designed to span nucleotide positions 2058 and 2059 in 23S rRNA. A PNA clamping probe was also designed to be complementary to the wild-type sequence (A2058\/A2059) and positioned to interfere with both the annealing of the 3\u2032 end of the backward inner primer and the concomitant extension. Prior to the LAMP assay, swab samples from suspected syphilitic lesions were boiled for DNA extraction.<\/jats:p>\n               <jats:p>\n                  <jats:bold>Results.<\/jats:bold> The assay had an equivalent detection limit of 1.0\u00d710<jats:sup>1<\/jats:sup> copies\/reaction and showed specificity against 38 pathogens. In the presence of a 4\u2009\u00b5M PNA probe, LAMP amplified up to 1.0\u00d7101 copies\/reaction using plasmids harbouring the complementary mutant sequences (A2058G or A2059G), whereas amplification was completely blocked for the wild-type sequence up to a concentration of 1.0\u00d710<jats:sup>3<\/jats:sup> copies\/reaction. For the 66 PCR-positive clinical specimens, the overall detection rate via LAMP was 93.9\u200a% (62\/66). Amplification was successful for all 53 mutant samples and was incomplete for all nine\u2009WT samples by the PNA-mediated LAMP assays.<\/jats:p>\n               <jats:p>\n                  <jats:bold>Conclusion.<\/jats:bold> We developed a PNA-mediated LAMP method that enabled us to rapidly identify <jats:italic>\n                     <jats:named-content content-type=\"species\">\n                        <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" ext-link-type=\"uri\" xlink:href=\"http:\/\/doi.org\/10.1601\/nm.7858\" xlink:type=\"simple\">T. pallidum<\/jats:ext-link>\n                     <\/jats:named-content>\n                  <\/jats:italic> and determine its macrolide susceptibility via a culture-independent protocol.<\/jats:p>","DOI":"10.1099\/jmm.0.001275","type":"journal-article","created":{"date-parts":[[2020,11,12]],"date-time":"2020-11-12T17:27:40Z","timestamp":1605202060000},"page":"1339-1345","source":"Crossref","is-referenced-by-count":7,"title":["A novel peptide nucleic acid- and loop-mediated isothermal amplification assay for the detection of mutations in the 23S rRNA gene of Treponema pallidum"],"prefix":"10.1099","volume":"69","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5866-8877","authenticated-orcid":false,"given":"Norihito","family":"Tarumoto","sequence":"first","affiliation":[{"name":"Department of Infectious Disease and Infection Control, Saitama Medical University, 38 Morohongo, Moroyama-machi, Iruma-gun, Saitama 350-0495, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kazuo","family":"Imai","sequence":"additional","affiliation":[{"name":"Department of Infectious Disease and Infection Control, Saitama Medical University, 38 Morohongo, Moroyama-machi, Iruma-gun, Saitama 350-0495, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shu-ichi","family":"Nakayama","sequence":"additional","affiliation":[{"name":"Department of Bacteriology I, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, Tokyo 162-8640, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ichiro","family":"Itoda","sequence":"additional","affiliation":[{"name":"Shirakaba Clinic, B-STEP 2F, 8-28, Sumiyoshi-cho, Shinjuku-ku, Tokyo 162-0065, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jun","family":"Sakai","sequence":"additional","affiliation":[{"name":"Department of Infectious Disease and Infection Control, Saitama Medical University, 38 Morohongo, Moroyama-machi, Iruma-gun, Saitama 350-0495, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takashi","family":"Murakami","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Saitama Medical University, 38 Morohongo, Moroyama-machi, Iruma-gun, Saitama 350-0495, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shigefumi","family":"Maesaki","sequence":"additional","affiliation":[{"name":"Department of Infectious Disease and Infection Control, Saitama Medical University, 38 Morohongo, Moroyama-machi, Iruma-gun, Saitama 350-0495, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Satoshi","family":"Hayakawa","sequence":"additional","affiliation":[{"name":"Division of Microbiology, Department of Pathology and Microbiology, Nihon University School of Medicine, 30-1, Oyaguchi Kami-cho, Itabashi-ku, Tokyo 173-8610, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Makoto","family":"Ohnishi","sequence":"additional","affiliation":[{"name":"Department of Bacteriology I, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, Tokyo 162-8640, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0912-4643","authenticated-orcid":false,"given":"Takuya","family":"Maeda","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory, Saitama Medical University, 38 Morohongo, Moroyama-machi, Iruma-gun, Saitama 350-0495, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"345","reference":[{"key":"R1","doi-asserted-by":"publisher","DOI":"10.1086\/429626"},{"key":"R2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0159309"},{"key":"R3","doi-asserted-by":"publisher","DOI":"10.7448\/IAS.19.1.20965"},{"key":"R4","doi-asserted-by":"publisher","DOI":"10.1097\/OLQ.0000000000000792"},{"key":"R5","doi-asserted-by":"publisher","DOI":"10.1097\/OLQ.0000000000000768"},{"key":"R6","doi-asserted-by":"publisher","DOI":"10.1038\/nmicrobiol.2016.245"},{"key":"R7","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijmm.2014.04.007"},{"key":"R8","doi-asserted-by":"publisher","DOI":"10.1017\/S0950268814002830"},{"key":"R9","doi-asserted-by":"publisher","DOI":"10.15698\/mic2016.09.523"},{"key":"R10","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6968.2008.01145.x"},{"key":"R11","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.45.1.1-12.2001"},{"key":"R12","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa040216"},{"key":"R13","doi-asserted-by":"publisher","DOI":"10.1099\/jmm.0.007542-0"},{"key":"R14","doi-asserted-by":"publisher","DOI":"10.1086\/498899"},{"key":"R15","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.44.3.806-807.2000"},{"key":"R16","doi-asserted-by":"publisher","DOI":"10.1097\/OLQ.0b013e31826f36de"},{"key":"R17","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.01167-18"},{"key":"R18","doi-asserted-by":"publisher","DOI":"10.1016\/j.mimet.2017.08.009"},{"key":"R19","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.39.5.1941-1946.2001"},{"key":"R20","doi-asserted-by":"publisher","DOI":"10.1111\/jdv.12734"},{"key":"R21","doi-asserted-by":"publisher","DOI":"10.1177\/0956462415624059"},{"key":"R22","first-page":"1","article-title":"Sexually transmitted diseases treatment guidelines, 2015","volume":"64","author":"Workowski","year":"2015","journal-title":"MMWR Recomm Rep"},{"key":"R23","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-11216-7"},{"key":"R24","doi-asserted-by":"publisher","DOI":"10.1007\/s10517-017-3925-5"},{"key":"R25","doi-asserted-by":"publisher","DOI":"10.1186\/s13071-016-1949-8"},{"key":"R26","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0151654"},{"key":"R27","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.02117-06"},{"key":"R28","article-title":"The use of a commercial loop-mediated isothermal amplification assay (TB-LAMP) for the detection of tuberculosis. http:\/\/apps.who.int\/iris\/bitstream\/10665\/83142\/1\/WHO_HTM_TB_2013.05_eng.pdf?ua=1 [accessed Feb 3 2019]","year":"2013"}],"container-title":["Journal of Medical Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.microbiologyresearch.org\/content\/journal\/jmm\/10.1099\/jmm.0.001275?crawler=true","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2020,12,21]],"date-time":"2020-12-21T17:12:36Z","timestamp":1608570756000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.microbiologyresearch.org\/content\/journal\/jmm\/10.1099\/jmm.0.001275"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,12,1]]},"references-count":28,"journal-issue":{"issue":"12","published-print":{"date-parts":[[2020,12,1]]}},"URL":"https:\/\/doi.org\/10.1099\/jmm.0.001275","relation":{},"ISSN":["0022-2615","1473-5644"],"issn-type":[{"value":"0022-2615","type":"print"},{"value":"1473-5644","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,12,1]]}}}