{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,24]],"date-time":"2026-08-24T06:19:17Z","timestamp":1787552357600,"version":"build-2736575974"},"reference-count":46,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,8,24]],"date-time":"2026-08-24T00:00:00Z","timestamp":1787529600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:p>\n                    <jats:italic>Streptococcus agalactiae<\/jats:italic>\n                    (Group B\n                    <jats:italic>Streptococcus<\/jats:italic>\n                    , GBS) is a major cause of neonatal and invasive infections, necessitating rapid and sensitive diagnostic approaches beyond conventional culture and PCR. Here, we developed an isothermal recombinase polymerase amplification (RPA)-MbCas12a CRISPR assay for fluorescence-based detection of GBS using a recently characterized MbCas12a variant from\n                    <jats:italic>Moraxella bovoculi<\/jats:italic>\n                    . The assay targets two conserved housekeeping genes,\n                    <jats:italic>sodA<\/jats:italic>\n                    and\n                    <jats:italic>groL<\/jats:italic>\n                    , and operates under isothermal conditions at 37\u202f\u00b0C with minimal sample processing. Analytical sensitivity reached 1.6\u202f\u00d7\u202f10\n                    <jats:sup>1<\/jats:sup>\n                    to 2.5\u202f\u00d7\u202f10\n                    <jats:sup>1<\/jats:sup>\n                    copies per reaction, while analytical specificity was confirmed against non-target bacterial species. The assay successfully detected all 14 tested GBS isolates directly from crude heat lysates and generated results within 1\u202fh. By combining rapid isothermal amplification, simplified sample preparation, and MbCas12a-mediated detection, this platform represents a promising approach for point-of-care GBS screening and infection control in resource-limited settings.\n                  <\/jats:p>","DOI":"10.3389\/fmicb.2026.1918484","type":"journal-article","created":{"date-parts":[[2026,8,24]],"date-time":"2026-08-24T05:28:04Z","timestamp":1787549284000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Rapid detection of Streptococcus agalactiae using an RPA-MbCas12a CRISPR assay targeting novel sodA and groL markers"],"prefix":"10.3389","volume":"17","author":[{"given":"Aldan","family":"Shamukhan","sequence":"first","affiliation":[{"name":"National Center for Biotechnology","place":["Astana, Kazakhstan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aisha","family":"Shaizadinova","sequence":"additional","affiliation":[{"name":"National Center for Biotechnology","place":["Astana, Kazakhstan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meruyert","family":"Amanzholova","sequence":"additional","affiliation":[{"name":"National Center for Biotechnology","place":["Astana, Kazakhstan"]},{"name":"Faculty of Biology and Biotechnology, Al-Farabi Kazakh National University","place":["Almaty, Kazakhstan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pavel","family":"Tarlykov","sequence":"additional","affiliation":[{"name":"National Center for Biotechnology","place":["Astana, Kazakhstan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sailau","family":"Abeldenov","sequence":"additional","affiliation":[{"name":"National Center for Biotechnology","place":["Astana, Kazakhstan"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,8,24]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"1371","DOI":"10.3390\/diagnostics16091371","article-title":"Integrated RPA\u2013CRISPR\/Cas12a Technology for Rapid Detection of Salmonella enterica","volume":"16","author":"Akimbekova","year":"2026","journal-title":"Diagnostics"},{"key":"ref2","doi-asserted-by":"publisher","first-page":"2914","DOI":"10.1016\/j.jmb.2019.01.035","article-title":"The double life of group B Streptococcus: asymptomatic colonizer and potent pathogen","volume":"431","author":"Armistead","year":"2019","journal-title":"J. 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Microbiol."},{"key":"ref5","doi-asserted-by":"publisher","first-page":"436","DOI":"10.1126\/science.aar6245","article-title":"CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity","volume":"360","author":"Chen","year":"2018","journal-title":"Science (New York, N.Y.)"},{"key":"ref6","doi-asserted-by":"publisher","first-page":"1926","DOI":"10.3389\/fmicb.2020.01926","article-title":"Highly sensitive and rapid identification of Streptococcus agalactiae based on multiple cross displacement amplification coupled with lateral flow biosensor assay","volume":"11","author":"Cheng","year":"2020","journal-title":"Front. Microbiol."},{"key":"ref7","doi-asserted-by":"publisher","first-page":"63","DOI":"10.1542\/pir.2023-006154","article-title":"Neonatal group B Streptococcus disease","volume":"45","author":"Coggins","year":"2024","journal-title":"Pediatr. Rev."},{"key":"ref8","doi-asserted-by":"publisher","first-page":"521","DOI":"10.1016\/j.jinf.2019.11.008","article-title":"Hospital clusters of invasive group B streptococcal disease: a systematic review","volume":"79","author":"Collin","year":"2019","journal-title":"J. Infect."},{"key":"ref9","doi-asserted-by":"crossref","first-page":"428","DOI":"10.1186\/1471-2334-14-428","article-title":"Emerging trends in invasive and noninvasive isolates of Streptococcus agalactiae in a Latin American hospital: a 17-year study","volume":"14","author":"Crespo-Ortiz","year":"2014","journal-title":"BMC Infect. Dis."},{"key":"ref10","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1186\/1471-2180-13-41","article-title":"Human Streptococcus agalactiae strains in aquatic mammals and fish","volume":"13","author":"Delannoy","year":"2013","journal-title":"BMC Microbiol."},{"key":"ref11","doi-asserted-by":"crossref","first-page":"947","DOI":"10.1111\/apa.17095","article-title":"Outbreak of group B Streptococcus in a neonatal care unit confirmed by whole-genome sequencing","volume":"113","author":"Delettre","year":"2024","journal-title":"Acta Paediatr (Oslo, Norway, 1992)"},{"key":"ref12","doi-asserted-by":"publisher","first-page":"417","DOI":"10.1086\/600303","article-title":"Diagnostic accuracy of a rapid real-time polymerase chain reaction assay for universal intrapartum group B streptococcus screening","volume":"49","author":"El Helali","year":"2009","journal-title":"Clin. Infect. 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Eng."},{"key":"ref20","doi-asserted-by":"publisher","first-page":"18","DOI":"10.1128\/AEM.00859-18","article-title":"Microevolution of Streptococcus agalactiae ST-261 from Australia indicates dissemination via imported Tilapia and ongoing adaptation to marine hosts or environment","volume":"84","author":"Kawasaki","year":"2018","journal-title":"Appl. Environ. Microbiol."},{"key":"ref21","first-page":"429","article-title":"Streptococcus agalactiae mastitis: a review","volume":"38","author":"Keefe","year":"1997","journal-title":"Can. Vet. J."},{"key":"ref22","doi-asserted-by":"publisher","first-page":"e0325724","DOI":"10.1128\/spectrum.03257-24","article-title":"CAMP-negative Streptococcus agalactiae strains exhibited complete or partial chromosomal deletions of the CAMP-factor encoding gene cfb","volume":"13","author":"Lai","year":"2025","journal-title":"Microbiol. Spectrum"},{"key":"ref23","doi-asserted-by":"crossref","first-page":"892","DOI":"10.1111\/apa.17152","article-title":"Maternal and neonatal group B Streptococcus colonisation: a systematic review and the meta-analysis of matched-pair studies","volume":"113","author":"Lee","year":"2024","journal-title":"Acta Paediatr (Oslo, Norway 1992)"},{"key":"ref24","doi-asserted-by":"publisher","first-page":"666","DOI":"10.1111\/jam.13925","article-title":"Evaluation of PCR primers targeting the groEL gene for the specific detection of Streptococcus agalactiae in the context of aquaculture","volume":"125","author":"Leigh","year":"2018","journal-title":"J. Appl. Microbiol."},{"key":"ref25","doi-asserted-by":"publisher","first-page":"344390","DOI":"10.1016\/j.aca.2025.344390","article-title":"CRISPR-based sensing platform for the group B streptococcus screening in pregnant women","volume":"1370","author":"Li","year":"2025","journal-title":"Anal. Chim. Acta"},{"key":"ref26","doi-asserted-by":"publisher","first-page":"149","DOI":"10.3201\/eid3101.231627","article-title":"Invasive group B Streptococcus infections caused by Hypervirulent clone of S. agalactiae sequence type 283, Hong Kong, China, 2021(1)","volume":"31","author":"Li","year":"2025","journal-title":"Emerg. Infect. Dis."},{"key":"ref27","doi-asserted-by":"publisher","first-page":"119906","DOI":"10.1016\/j.cca.2024.119906","article-title":"Establishment of a rapid detection method for Mycoplasma pneumoniae based on RPA-CRISPR-Cas12a technology","volume":"564","author":"Li","year":"2025","journal-title":"Clin Chim Acta"},{"key":"ref28","doi-asserted-by":"publisher","first-page":"e49303","DOI":"10.1097\/MD.0000000000049303","article-title":"Development of a one-pot RPA-CRISPR\/Cas12a assay for rapid multiplex detection of respiratory pathogens","volume":"105","author":"Liao","year":"2026","journal-title":"Medicine"},{"key":"ref29","doi-asserted-by":"publisher","first-page":"775","DOI":"10.1186\/1471-2164-14-775","article-title":"Comparative genomics analysis of Streptococcus agalactiae reveals that isolates from cultured tilapia in China are closely related to the human strain A909","volume":"14","author":"Liu","year":"2013","journal-title":"BMC Genomics"},{"key":"ref30","doi-asserted-by":"publisher","first-page":"19","DOI":"10.1016\/j.trac.2017.10.015","article-title":"Recombinase polymerase amplification: basics, applications and recent advances","volume":"98","author":"Lobato","year":"2018","journal-title":"Trends Anal Chem"},{"key":"ref31","doi-asserted-by":"crossref","first-page":"S160","DOI":"10.1093\/cid\/cix656","article-title":"Infant group B streptococcal disease incidence and serotypes worldwide: systematic review and meta-analyses","volume":"65","author":"Madrid","year":"2017","journal-title":"Clin. Infect. Dis."},{"key":"ref32","doi-asserted-by":"publisher","first-page":"117189","DOI":"10.1016\/j.diagmicrobio.2025.117189","article-title":"Investigation of Streptococcus agalactiae colonisation in pregnant women using culture and a novel qPCR kit","volume":"114","author":"Mazlumo\u011flu","year":"2026","journal-title":"Diagn. Microbiol. Infect. Dis."},{"key":"ref33","first-page":"1236","article-title":"Epidemiology of antimicrobial resistance genes in Streptococcus agalactiae sequences from a public database in a one health perspective","volume":"11","author":"Meroni","year":"2022","journal-title":"Antibiotics (Basel, Switzerland)"},{"key":"ref34","doi-asserted-by":"publisher","first-page":"2721","DOI":"10.3201\/eid2611.201669","article-title":"Multidrug-resistant Hypervirulent group B Streptococcus in neonatal invasive infections, France, 2007-2019","volume":"26","author":"Plainvert","year":"2020","journal-title":"Emerg. Infect. Dis."},{"key":"ref35","doi-asserted-by":"publisher","first-page":"41","DOI":"10.1128\/JCM.36.1.41-47.1998","article-title":"Identification of streptococci to species level by sequencing the gene encoding the manganese-dependent superoxide dismutase","volume":"36","author":"Poyart","year":"1998","journal-title":"J. Clin. Microbiol."},{"key":"ref36","doi-asserted-by":"publisher","first-page":"18","DOI":"10.1128\/microbiolspec.GPP3-0007-2018","article-title":"Group B Streptococcus (Streptococcus agalactiae)","volume":"7","author":"Raabe","year":"2019","journal-title":"Microbiol. Spectrum"},{"key":"ref37","doi-asserted-by":"publisher","first-page":"2590","DOI":"10.1128\/JCM.00582-17","article-title":"Reliable detection of group B Streptococcus in the clinical laboratory","volume":"55","author":"Rosa-Fraile","year":"2017","journal-title":"J. Clin. 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Res."},{"key":"ref41","doi-asserted-by":"publisher","first-page":"120281","DOI":"10.1016\/j.cca.2025.120281","article-title":"Rapid detection of Klebsiella pneumoniae based on one-tube RPA-CRISPR\/Cas12a system","volume":"573","author":"Wu","year":"2025","journal-title":"Clin. Chim. Acta"},{"key":"ref42","doi-asserted-by":"publisher","first-page":"8","DOI":"10.1186\/s12941-023-00558-2","article-title":"CRISPR\/Cas12a-based assay for the rapid and high-sensitivity detection of Streptococcus agalactiae colonization in pregnant women with premature rupture of membrane","volume":"22","author":"Yu","year":"2023","journal-title":"Ann. Clin. Microbiol. Antimicrob."},{"key":"ref43","doi-asserted-by":"publisher","first-page":"e0081925","DOI":"10.1128\/jcm.00819-25","article-title":"An extraction-free and one-pot two-temperature CRISPR\/Cas12b system for visual detection of group B Streptococcus by integrating with RPA","volume":"63","author":"Zhao","year":"2025","journal-title":"J. Clin. Microbiol."},{"key":"ref44","doi-asserted-by":"publisher","first-page":"581","DOI":"10.2478\/jvetres-2022-0069","article-title":"Antimicrobial resistance and virulence genes of Streptococcus agalactiae isolated from mastitis Milk samples in China","volume":"66","author":"Zhao","year":"2022","journal-title":"J Vet Res"},{"key":"ref45","doi-asserted-by":"publisher","first-page":"126574","DOI":"10.1016\/j.talanta.2024.126574","article-title":"Fast, portable and sensitive detection of group B streptococcus DNA using one-pot MIRA-CRISPR system with suboptimal PAM","volume":"279","author":"Zheng","year":"2024","journal-title":"Talanta"},{"key":"ref46","doi-asserted-by":"publisher","first-page":"1189093","DOI":"10.3389\/fmicb.2023.1189093","article-title":"Analysis of molecular characteristics of CAMP-negative Streptococcus agalactiae strains","volume":"14","author":"Zhou","year":"2023","journal-title":"Front. Microbiol."}],"container-title":["Frontiers in Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2026.1918484\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,24]],"date-time":"2026-08-24T05:28:06Z","timestamp":1787549286000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2026.1918484\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,24]]},"references-count":46,"alternative-id":["10.3389\/fmicb.2026.1918484"],"URL":"https:\/\/doi.org\/10.3389\/fmicb.2026.1918484","relation":{},"ISSN":["1664-302X"],"issn-type":[{"value":"1664-302X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,24]]},"article-number":"1918484"}}