{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,18]],"date-time":"2026-10-18T13:19:48Z","timestamp":1792329588001,"version":"4.4.0"},"reference-count":30,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,8,8]],"date-time":"2024-08-08T00:00:00Z","timestamp":1723075200000},"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. Cell. Infect. Microbiol."],"abstract":"<jats:p>\n                    <jats:italic>Mycoplasma pneumoniae<\/jats:italic>\n                    is a significant pathogen responsible for community-acquired pneumonia, predominantly affecting children and adolescents. Here, we devised a rapid method for\n                    <jats:italic>M. pneumoniae<\/jats:italic>\n                    that combined multiple cross displacement amplification (MCDA) with real-time fluorescence technology. A set of ten primers, which were specifically designed for\n                    <jats:italic>M. pneumoniae<\/jats:italic>\n                    detection, were employed in a real-time fluorescence MCDA reaction. Of these, one primer incorporated a restriction endonuclease recognition sequence, a fluorophore, and a quencher, facilitating real-time fluorescence detection. The real-time (RT)-MCDA reactions were monitored in a simple real-time fluorescence instrument and conducted under optimised conditions (64\u00b0C for 40 min). The detection limit of the\n                    <jats:italic>M. pneumoniae<\/jats:italic>\n                    RT-MCDA assay for genomic DNA extracted from\n                    <jats:italic>M. pneumoniae<\/jats:italic>\n                    culture was down to 43 fg\/\u00b5l. This assay accurately identified\n                    <jats:italic>M. pneumoniae<\/jats:italic>\n                    strains without cross-reacting with other bacteria. To validate its practical application, we tested the\n                    <jats:italic>M. pneumoniae<\/jats:italic>\n                    RT-MCDA assay using genomic DNA extracted from clinical samples. The assay\u2019s detection capability proved comparable with real-time PCR, MCDA-based biosensor detection, and visual inspection under blue light. The entire process, including rapid DNA extraction and real-time MCDA detection, was completed within 1 h. Overall, the\n                    <jats:italic>M. pneumoniae<\/jats:italic>\n                    RT-MCDA assay reported here is a simple and effective diagnostic tool for rapid\n                    <jats:italic>M. pneumoniae<\/jats:italic>\n                    detection, which holds significant potential for point-of-care testing and in resource-limited regions.\n                  <\/jats:p>","DOI":"10.3389\/fcimb.2024.1423155","type":"journal-article","created":{"date-parts":[[2024,8,8]],"date-time":"2024-08-08T01:10:33Z","timestamp":1723079433000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":12,"title":["Real-time fluorescent multiple cross displacement amplification for rapid and sensitive Mycoplasma pneumoniae detection"],"prefix":"10.3389","volume":"14","author":[{"given":"Fei","family":"Xiao","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenjian","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jin","family":"Fu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaolan","family":"Huang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nan","family":"Jia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunrong","family":"Sun","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zheng","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Baoying","family":"Zheng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juan","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yi","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lihui","family":"Meng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,8,8]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"677","DOI":"10.3390\/bios12090677","article-title":"Isothermal amplification technology for disease diagnosis","volume":"12","author":"Boonbanjong","year":"2022","journal-title":"Biosensors (Basel)"},{"key":"B2","doi-asserted-by":"publisher","first-page":"137","DOI":"10.1016\/j.jmii.2013.03.015","article-title":"Comparison of real-time polymerase chain reaction and serological tests for the confirmation of Mycoplasma pneumoniae infection in children with clinical diagnosis of atypical pneumonia","volume":"47","author":"Chang","year":"2014","journal-title":"J. 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