{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T13:52:02Z","timestamp":1791553922085,"version":"4.3.4"},"reference-count":42,"publisher":"American Chemical Society (ACS)","issue":"10","license":[{"start":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T00:00:00Z","timestamp":1790640000000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T00:00:00Z","timestamp":1790640000000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T00:00:00Z","timestamp":1790640000000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/na","name":"Russian state-funded project for ICBFM SB RAS","doi-asserted-by":"publisher","award":["125012300657-2"],"award-info":[{"award-number":["125012300657-2"]}],"id":[{"id":"10.13039\/na","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,10,9]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Reverse transcriptases from bacterial group II introns (GII RTs) demonstrate superior thermostability and processivity compared with retroviral counterparts. While GII RTs have proven advantageous for RNA-seq, their performance in other applications, such as reverse transcription loop-mediated isothermal amplification (RT-LAMP), remains poorly studied. RT-LAMP is a widely used method suitable for point-of-care diagnostics of RNA viruses and other pathogens. In this study, we compared the performance in RT-LAMP of GII RT from the thermophilic bacterium Anoxybacillus flavithermus (Afl RT) with that of the retroviral RNase H-deficient reverse transcriptase M-MuLV. A duplex RT-LAMP targeting SARS-CoV-2 and MS2 phage genomic RNA (internal control) served as a model. Preheating at 95 \u00b0C and 1.5 M betaine increased the efficiency of RT-LAMP with both reverse transcriptases. Under these conditions, the optimal temperature range and time of reverse transcription were similar for both enzymes, 55\u201360 \u00b0C and 15 min, respectively. However, high concentrations of Afl RT (&amp;gt;50 U\/reaction) inhibited RT-LAMP. Notably, RT-LAMP with Afl RT demonstrated higher sensitivity for detecting both in vitro-transcribed RNA fragments and SARS-CoV-2\u2013positive samples. In validation against RT-qPCR using nasal swabs (200 SARS-CoV-2-positive and 200 negative samples) from patients with respiratory symptoms, the observed sensitivity of RT-LAMP with Afl RT was 100% (200\/200; 95% CI, 98.17\u2013100%), whereas with M-MuLV RT, sensitivity was 97.0% (194\/200; 95% CI, 93.58\u201398.89%). Specificity was 100% for both RTs (200\/200; 95% CI, 98.17\u2013100%). These observations demonstrate that Afl RT is suitable for RT-LAMP and may increase the sensitivity of LAMP-based diagnostics.<\/jats:p>","DOI":"10.1021\/acsinfecdis.6c00293","type":"journal-article","created":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T20:02:09Z","timestamp":1790712129000},"page":"3351-3361","source":"Crossref","is-referenced-by-count":0,"title":["Performance\nof a Group II Intron Reverse Transcriptase\nfrom\n                    <i>Anoxybacillus flavithermus<\/i>\n                    in\nDuplex RT-LAMP for SARS-CoV-2 Detection"],"prefix":"10.1021","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3754-5824","authenticated-orcid":true,"given":"Igor P.","family":"Oscorbin","sequence":"first","affiliation":[{"name":"Siberian Branch of the Russian Academy of Sciences (ICBFM SB RAS) , , 8, Lavrentiev Avenue , ,","place":["Novosibirsk, Russia, 630090"]},{"name":"Novosibirsk State University , , 1, Pirogova Street , ,","place":["Novosibirsk, Russia, 630090"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elizaveta D.","family":"Baranova","sequence":"additional","affiliation":[{"name":"Siberian Branch of the Russian Academy of Sciences (ICBFM SB RAS) , , 8, Lavrentiev Avenue , ,","place":["Novosibirsk, Russia, 630090"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maxim L.","family":"Filipenko","sequence":"additional","affiliation":[{"name":"Siberian Branch of the Russian Academy of Sciences (ICBFM SB RAS) , , 8, Lavrentiev Avenue , ,","place":["Novosibirsk, Russia, 630090"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,29]]},"reference":[{"issue":"5312","key":"2026100909094312000_cit1","doi-asserted-by":"publisher","first-page":"561","DOI":"10.1126\/science.276.5312.561","article-title":"Reverse Transcriptase Motifs in the Catalytic Subunit\nof Telomerase","volume":"276","author":"Lingner","year":"1997","journal-title":"Science"},{"key":"2026100909094312000_cit2","doi-asserted-by":"publisher","first-page":"153","DOI":"10.1016\/j.virusres.2016.12.019","article-title":"Viral Reverse\nTranscriptases","volume":"234","author":"Mene\u0301ndez-Arias","year":"2017","journal-title":"Virus Res."},{"issue":"10","key":"2026100909094312000_cit3","doi-asserted-by":"publisher","first-page":"657","DOI":"10.1093\/protein\/gzs034","article-title":"Generation\nand Characterization of\nNew Highly Thermostable and Processive M-MuLV Reverse Transcriptase\nVariants","volume":"25","author":"Baranauskas","year":"2012","journal-title":"Protein Eng., Des. Sel."},{"issue":"2","key":"2026100909094312000_cit4","doi-asserted-by":"publisher","first-page":"473","DOI":"10.1093\/nar\/gkn952","article-title":"Novel Mutations in Moloney Murine Leukemia Virus Reverse\nTranscriptase Increase Thermostability through Tighter Binding to\nTemplate-Primer","volume":"37","author":"Arezi","year":"2009","journal-title":"Nucleic Acids Res."},{"issue":"8","key":"2026100909094312000_cit5","doi-asserted-by":"publisher","first-page":"551","DOI":"10.1093\/protein\/gzx046","article-title":"Further Increase in\nThermostability of Moloney Murine\nLeukemia Virus Reverse Transcriptase by Mutational Combination","volume":"30","author":"Baba","year":"2017","journal-title":"Protein Eng., Des. Sel."},{"issue":"7","key":"2026100909094312000_cit6","doi-asserted-by":"publisher","first-page":"1209","DOI":"10.1007\/s10529-012-0904-9","article-title":"Improving\nthe Thermal Stability of\nAvian Myeloblastosis Virus Reverse Transcriptase \u03b1-Subunit by\nSite-Directed Mutagenesis","volume":"34","author":"Konishi","year":"2012","journal-title":"Biotechnol. Lett"},{"issue":"12","key":"2026100909094312000_cit7","doi-asserted-by":"publisher","first-page":"2339","DOI":"10.1080\/09168451.2017.1394790","article-title":"Generation of Thermostable Moloney\nMurine Leukemia Virus Reverse Transcriptase Variants Using Site Saturation\nMutagenesis Library and Cell-Free Protein Expression System","volume":"81","author":"Katano","year":"2017","journal-title":"Biosci., Biotechnol., Biochem."},{"issue":"3","key":"2026100909094312000_cit8","doi-asserted-by":"publisher","first-page":"299","DOI":"10.1016\/j.jbiotec.2010.09.961","article-title":"Increase in Thermal\nStability of Moloney Murine Leukaemia Virus Reverse Transcriptase\nby Site-Directed Mutagenesis","volume":"150","author":"Yasukawa","year":"2010","journal-title":"J. Biotechnol."},{"issue":"2","key":"2026100909094312000_cit9","doi-asserted-by":"publisher","first-page":"301","DOI":"10.1016\/j.ab.2010.01.024","article-title":"Mutant of Moloney Murine Leukemia\nVirus Reverse Transcriptase Exhibits Higher Resistance to Common RT-QPCR\nInhibitors","volume":"400","author":"Arezi","year":"2010","journal-title":"Anal. 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Environ. Microbiol."},{"issue":"28","key":"2026100909094312000_cit17","doi-asserted-by":"publisher","first-page":"9069","DOI":"10.1021\/bi982799l","article-title":"RNA and\nProtein Catalysis in Group II Intron Splicing\nand Mobility Reactions Using Purified Components \u2020","volume":"38","author":"Saldanha","year":"1999","journal-title":"Biochemistry"},{"issue":"5","key":"2026100909094312000_cit18","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkab1207","article-title":"Identification and\nClassification of Reverse Transcriptases in Bacterial\nGenomes and Metagenomes","volume":"50","author":"Sharifi","year":"2022","journal-title":"Nucleic Acids Res."},{"issue":"21","key":"2026100909094312000_cit19","doi-asserted-by":"publisher","first-page":"2910","DOI":"10.1101\/gad.11.21.2910","article-title":"A Bacterial\nGroup II Intron Encoding Reverse Transcriptase,\nMaturase, and DNA Endonuclease Activities: Biochemical Demonstration\nof Maturase Activity and Insertion of New Genetic Information within\nthe Intron","volume":"11","author":"Matsuura","year":"1997","journal-title":"Genes Dev."},{"issue":"1","key":"2026100909094312000_cit20","doi-asserted-by":"publisher","first-page":"49","DOI":"10.3390\/biom14010049","article-title":"A Novel\nThermostable and Processive Reverse Transcriptase\nfrom a Group II Intron of Anoxybacillus Flavithermus","volume":"14","author":"Oscorbin","year":"2024","journal-title":"Biomolecules"},{"issue":"5","key":"2026100909094312000_cit21","doi-asserted-by":"publisher","first-page":"1014","DOI":"10.1111\/j.1365-2672.2012.05384.x","article-title":"PCR Inhibitors - Occurrence,\nProperties and Removal","volume":"113","author":"Schrader","year":"2012","journal-title":"J. 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Sci."},{"issue":"10","key":"2026100909094312000_cit35","doi-asserted-by":"publisher","first-page":"947","DOI":"10.1007\/s12033-015-9886-x","article-title":"Large Fragment of DNA Polymerase\nI from Geobacillus Sp. 777: Cloning and Comparison with DNA Polymerases\nI in Practical Applications","volume":"57","author":"Oscorbin","year":"2015","journal-title":"Mol. Biotechnol."},{"issue":"12","key":"2026100909094312000_cit36","doi-asserted-by":"publisher","first-page":"1809","DOI":"10.3390\/biology11121809","article-title":"Comparison of Reverse Transcriptase\n(RT) Activities of Various M-MuLV RTs for RT-LAMP Assays","volume":"11","author":"Oscorbin","year":"2022","journal-title":"Biology"},{"issue":"1","key":"2026100909094312000_cit37","doi-asserted-by":"publisher","first-page":"265","DOI":"10.1093\/nar\/16.1.265","article-title":"Isolation of Cloned\nMoloney Murine Leukemia Virus Reverse Transcriptase Lacking Ribonuclease\nH Activity","volume":"16","author":"Kotewicz","year":"1988","journal-title":"Nucleic Acids Res."},{"issue":"1","key":"2026100909094312000_cit38","doi-asserted-by":"publisher","first-page":"488","DOI":"10.1093\/nar\/gkac1221","article-title":"Kinetics\nof Elementary Steps in Loop-Mediated Isothermal Amplification (LAMP)\nShow That Strand Invasion during Initiation Is Rate-Limiting","volume":"51","author":"Dangerfield","year":"2023","journal-title":"Nucleic Acids Res."},{"issue":"43","key":"2026100909094312000_cit39","doi-asserted-by":"publisher","first-page":"13804","DOI":"10.1021\/jacs.5b08144","article-title":"Innate Reverse Transcriptase\nActivity of DNA Polymerase for Isothermal RNA Direct Detection","volume":"137","author":"Shi","year":"2015","journal-title":"J. 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Microbiol."}],"container-title":["ACS Infectious Diseases"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acsinfecdis.6c00293","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/aidcbc\/article-pdf\/12\/10\/3351\/68702903\/acsinfecdis.6c00293.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/aidcbc\/article-pdf\/12\/10\/3351\/68702903\/acsinfecdis.6c00293.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T13:23:21Z","timestamp":1791552201000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/aidcbc\/article\/12\/10\/3351\/5441842\/Performance-of-a-Group-II-Intron-Reverse"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,29]]},"references-count":42,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2026,9,29]]},"published-print":{"date-parts":[[2026,10,9]]}},"URL":"https:\/\/doi.org\/10.1021\/acsinfecdis.6c00293","relation":{},"ISSN":["2373-8227"],"issn-type":[{"value":"2373-8227","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,29]]}}}