{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,22]],"date-time":"2026-07-22T06:07:21Z","timestamp":1784700441547,"version":"3.55.0"},"reference-count":24,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,7,22]],"date-time":"2026-07-22T00:00:00Z","timestamp":1784678400000},"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:sec>\n                    <jats:title>Objective<\/jats:title>\n                    <jats:p>\n                      <jats:italic>Pseudomonas aeruginosa (P. aeruginosa)<\/jats:italic>\n                      is a common contaminant in traditional Chinese medicine (TCM) herbal pieces, and its detection is often hindered by polysaccharides, polyphenols, and other inhibitory compounds in the herbal matrix. This study aimed to develop a rapid, sensitive, and matrix-tolerant method for detecting P. aeruginosa in TCM herbal pieces for routine quality control.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      A loop-mediated isothermal amplification (LAMP) assay targeting the species-specific\n                      <jats:italic>ecfX<\/jats:italic>\n                      gene was developed and integrated with a fluorescent microsphere\u2013based lateral flow strip (LFS) for visual result interpretation. LAMP primers were dual labeled with digoxigenin and biotin to enable specific amplicon to capture at the test and control lines of the LFS. Reaction conditions for both LAMP reaction and LFS were optimized using genomic DNA extracted from\n                      <jats:italic>P. aeruginosa<\/jats:italic>\n                      -spiked powdered herbal samples. Analytical performance, including specificity, sensitivity, reproducibility, and limit of detection (LOD), was rigorously evaluated. The validated method was then applied to six commonly used TCM herbal pieces, including Astragali radix and Angelicae sinensis radix. Additionally, the assay performance was validated using six representative TCM herbal pieces and 90 clinical-relevant herbal samples in parallel with qPCR.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      The LAMP-LFS assay demonstrated strict specificity for\n                      <jats:italic>P. aeruginosa<\/jats:italic>\n                      , with no cross-reactivity observed against any non-target bacterial strains tested. Optimal LAMP conditions were determined as 65\u00a0\u00b0C for 60\u00a0min in the presence of 6 mM Mg\n                      <jats:sup>2+<\/jats:sup>\n                      and 1.4 mM dNTPs. The entire detection workflow, including DNA extraction, amplification, and strip readout, was completed within approximately 70\u00a0min. The assay achieved a limit of detection of 1 \u00d7 10\u00b2 CFU\/250 mL in spiked herbal matrices, with high reproducibility (CV &amp;lt; 5%). Consistent detection was observed across six different TCM herbal species, indicating strong tolerance to complex phytochemical backgrounds. In parallel validation with qPCR, the assay demonstrated a sensitivity of 94.0%, specificity of 97.5%, and overall accuracy of 95.6%, with a Cohen\u2019s kappa coefficient of 0.92.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      The developed fluorescent microsphere\u2013based LAMP-LFS assay provides a rapid, sensitive, and reproducible platform for detecting\n                      <jats:italic>P. aeruginosa<\/jats:italic>\n                      in TCM herbal pieces. Compared with conventional PCR-based approaches, this method demonstrates improved tolerance to inhibitory herbal matrices and enables reliable on-site applicability with minimal instrumentation requirements. The assay shows potential for routine microbiological quality control and field-based monitoring of TCM herbal materials.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fcimb.2026.1861254","type":"journal-article","created":{"date-parts":[[2026,7,22]],"date-time":"2026-07-22T05:56:18Z","timestamp":1784699778000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Rapid and sensitive detection of Pseudomonas aeruginosa in traditional Chinese medicine herbal pieces using loop-mediated isothermal amplification combined with fluorescent microsphere\u2013based lateral flow strip"],"prefix":"10.3389","volume":"16","author":[{"given":"Wenlin","family":"Tian","sequence":"first","affiliation":[{"name":"Nanobiosensing and Microfluidic Point-of-Care Testing, Key Laboratory of Luzhou, Department of Clinical Laboratory, The Affiliated Traditional Chinese Medicine Hospital, Southwest Medical University","place":["Luzhou, Sichuan, China"]},{"name":"Laboratory Department, Hejiang County Traditional Chinese Medicine Hospital","place":["Luzhou, Sichuan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chao","family":"Yang","sequence":"additional","affiliation":[{"name":"Laboratory Department, Hejiang County Traditional Chinese Medicine Hospital","place":["Luzhou, Sichuan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhilan","family":"Chen","sequence":"additional","affiliation":[{"name":"Laboratory Department, Hejiang County Traditional Chinese Medicine Hospital","place":["Luzhou, Sichuan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingtao","family":"Wen","sequence":"additional","affiliation":[{"name":"Laboratory Department, Hejiang County Traditional Chinese Medicine Hospital","place":["Luzhou, Sichuan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yujie","family":"Zhong","sequence":"additional","affiliation":[{"name":"Laboratory Department, Hejiang County Traditional Chinese Medicine Hospital","place":["Luzhou, Sichuan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yongcan","family":"Guo","sequence":"additional","affiliation":[{"name":"Nanobiosensing and Microfluidic Point-of-Care Testing, Key Laboratory of Luzhou, Department of Clinical Laboratory, The Affiliated Traditional Chinese Medicine Hospital, Southwest Medical University","place":["Luzhou, Sichuan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,7,22]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.3390\/bios13110988","article-title":"Detection of reverse transcriptase LAMP-amplified nucleic acid from oropharyngeal viral swab samples using biotinylated DNA probes through a lateral flow assay","volume":"13","author":"Agarwal","year":"2023","journal-title":"Biosens.-Basel"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.3389\/fpubh.2026.1798338","article-title":"LAMP-LFD: establishment and evaluation of a new diagnostic platform for SARS-CoV-2 rapid detection","volume":"14","author":"Diao","year":"2026","journal-title":"Front. Public Health"},{"key":"B3","doi-asserted-by":"publisher","first-page":"111","DOI":"10.1186\/s13756-024-01468-4","article-title":"Occurrence of chlorine-resistant Pseudomonas aeruginosa in hospital water systems: threat of waterborne infections for patients","volume":"13","author":"Gholipour","year":"2024","journal-title":"Antimicrobial Resistance Infection Control"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.3389\/fphar.2025.1658241","article-title":"Advances in research on novel technologies for the detection of exogenous contaminants in traditional Chinese medicine","volume":"16","author":"Guo","year":"2025","journal-title":"Front. Pharmacol."},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.3390\/bios15110750","article-title":"Rapid on-site detection of Pseudomonas aeruginosa via ecfX-targeted loop-mediated isothermal amplification","volume":"15","author":"He","year":"2025","journal-title":"Biosens.-Basel"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0302783","article-title":"Development of a multiplex loop-mediated isothermal amplification (LAMP) for the diagnosis of bacterial periprosthetic joint infection","volume":"19","author":"Jang","year":"2024","journal-title":"PLoS One"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1155\/2023\/9975620","article-title":"Simultaneous and visual detection of KPC and NDM carbapenemase-encoding genes using asymmetric PCR and multiplex lateral flow strip","volume":"2023","author":"Lai","year":"2023","journal-title":"J. Anal. Methods Chem."},{"key":"B8","doi-asserted-by":"publisher","first-page":"20","DOI":"10.1016\/j.mimet.2007.03.008","article-title":"Improved reliability of Pseudomonas aeruginosa PCR detection by the use of the species-specific ecfX gene target","volume":"70","author":"Lavenir","year":"2007","journal-title":"J. Microbiol. Methods"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.3390\/ijms26135933","article-title":"Optimization of loop-mediated isothermal amplification (LAMP) for the rapid detection of nosocomial pathogens on environmental surfaces","volume":"26","author":"Marino","year":"2025","journal-title":"Int. J. Mol. Sci."},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.3390\/foods11030322","article-title":"A review of isothermal amplification methods and food-origin inhibitors against detecting food-borne pathogens","volume":"11","author":"Moon","year":"2022","journal-title":"Foods"},{"key":"B11","doi-asserted-by":"publisher","first-page":"5916","DOI":"10.1038\/s41598-024-55241-z","article-title":"Evaluation of molecular inhibitors of loop-mediated isothermal amplification (LAMP)","volume":"14","author":"Nwe","year":"2024","journal-title":"Sci. Rep."},{"key":"B12","doi-asserted-by":"publisher","first-page":"4519","DOI":"10.1016\/j.csbj.2023.09.008","article-title":"Bst polymerase - a humble relative of Taq polymerase","volume":"21","author":"Oscorbin","year":"2023","journal-title":"Comput. Struct. Biotechnol. J."},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.3390\/bios12100857","article-title":"Principles and applications of loop-mediated isothermal amplification to point-of-care tests","volume":"12","author":"Park","year":"2022","journal-title":"Biosens.-Basel"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.01687-23","article-title":"Clinical validation of loop-mediated isothermal amplification for the detection of Escherichia coli sequence type complex 131","volume":"62","author":"Peirano","year":"2024","journal-title":"J. Clin. Microbiol."},{"key":"B15","doi-asserted-by":"publisher","first-page":"101572","DOI":"10.1016\/j.jpha.2026.101572","article-title":"Revolutionizing traditional Chinese medicine research: advancements in traditional machine learning and deep learning for AI-driven applications","volume":"61","author":"Ren","year":"2026","journal-title":"J. Pharm. Anal."},{"key":"B16","doi-asserted-by":"publisher","first-page":"2116","DOI":"10.21608\/nrmj.2023.317346","article-title":"Microbial contamination of cosmetics and the pharmaceutical products, and their preservation strategies: a comprehensive review","volume":"7","author":"Roy","year":"2023","journal-title":"Novel Res. Microbiol. J."},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1016\/j.forsciint.2023.111773","article-title":"PCR inhibitors and facilitators - their role in forensic DNA analysis","volume":"349","author":"Vajpayee","year":"2023","journal-title":"Forensic Sci. Int."},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2022.820431","article-title":"Pseudomonas aeruginosa detection using conventional PCR and quantitative real-time PCR based on species-specific novel gene targets identified by pangenome analysis","volume":"13","author":"Wang","year":"2022","journal-title":"Front. Microbiol."},{"key":"B19","doi-asserted-by":"publisher","first-page":"7137","DOI":"10.1038\/s41598-022-11320-7","article-title":"Investigation and validation of labelling loop mediated isothermal amplification (LAMP) products with different nucleotide modifications for various downstream analysis","volume":"12","author":"Warmt","year":"2022","journal-title":"Sci. Rep."},{"key":"B20","doi-asserted-by":"publisher","first-page":"21028","DOI":"10.1021\/acs.est.5c07943","article-title":"Perfluorooctanoic acid can increase biofilm resilience and virulence in drinking water distribution systems","volume":"59","author":"Wang","year":"2025","journal-title":"Environ. Sci. Technol."},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2021.698929","article-title":"A rapid and sensitive detection method for Pseudomonas aeruginosa using visualized recombinase polymerase amplification and lateral flow strip technology","volume":"11","author":"Yang","year":"2021","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.3390\/jof11060411","article-title":"Research progress and management strategies for the common mycotoxin contamination of traditional Chinese medicines","volume":"11","author":"Yang","year":"2025","journal-title":"J. Fungi"},{"key":"B23","doi-asserted-by":"publisher","first-page":"3605","DOI":"10.19540\/j.cnki.cjcmm.20250529.301","article-title":"Research progress in machine learning in processing and quality evaluation of traditional Chinese medicine decoction pieces","volume":"50","author":"Zhang","year":"2025","journal-title":"Zhongguo Zhong Yao Za Zhi"},{"key":"B24","doi-asserted-by":"publisher","first-page":"797","DOI":"10.1089\/jmf.2024.k.0004","article-title":"Rapid mold detection in Chinese herbal medicine using enhanced deep learning technology","volume":"27","author":"Zhu","year":"2024","journal-title":"J. Med. Food"}],"container-title":["Frontiers in Cellular and Infection Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2026.1861254\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,22]],"date-time":"2026-07-22T05:56:19Z","timestamp":1784699779000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2026.1861254\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,22]]},"references-count":24,"alternative-id":["10.3389\/fcimb.2026.1861254"],"URL":"https:\/\/doi.org\/10.3389\/fcimb.2026.1861254","relation":{},"ISSN":["2235-2988"],"issn-type":[{"value":"2235-2988","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,22]]},"article-number":"1861254"}}