{"doi":"10.3389/fchem.2021.815189","title":"Rapid Detection and Differentiation of Legionella pneumophila and Non-Legionella pneumophila Species by Using Recombinase Polymerase Amplification Combined With EuNPs-Based Lateral Flow Immunochromatography","abstract":"<jats:p><jats:italic>Legionella</jats:italic>, a waterborne pathogen, is the main cause of Legionnaires’ disease. Therefore, timely and accurate detection and differentiation of <jats:italic>Legionella pneumophila</jats:italic> and non<jats:italic>-Legionella pneumophila</jats:italic> species is crucial. In this study, we develop an easy and rapid recombinase polymerase amplification assay combined with EuNPs-based lateral flow immunochromatography (EuNPs-LFIC-RPA) to specifically distinguish <jats:italic>Legionella pneumophila</jats:italic> and non<jats:italic>-Legionella pneumophila</jats:italic>. We designed primers based on the <jats:italic>mip</jats:italic> gene of <jats:italic>Legionella pneumophila</jats:italic> and the 5S rRNA gene of non-<jats:italic>Legionella pneumophila.</jats:italic> The recombinase polymerase amplification reaction could go to completion in 10 min at 37°C, and the amplification products could be detected within 5 min with EuNPs-LFIC strips. Using a florescent test strip reader, the quantitative results were achieved by reading the colored signal intensities on the strips. The sensitivity was 1.6 × 10<jats:sup>1</jats:sup> CFU/ml, and a linear standard linear curve plotted from the test strip reader had a correlation coefficient for the determination of <jats:italic>Legionella pneumophila</jats:italic> (<jats:italic>R</jats:italic><jats:sup>2</jats:sup> = 0.9516). Completed concordance for the presence or absence of <jats:italic>Legionella pneumophila</jats:italic> by EuNPs-LFIC-RPA and qPCR was 97.32% (<jats:italic>κ</jats:italic> = 0.79, 95% CI), according to an analysis of practical water samples (<jats:italic>n</jats:italic> = 112). In short, this work shows the feasibility of EuNPs-LFIC-RPA for efficient and rapid monitoring of <jats:italic>Legionella pneumophila</jats:italic> and non-<jats:italic>Legionella pneumophila</jats:italic> in water samples.</jats:p>","journal":"Frontiers in Chemistry","year":2022,"id":644027,"datarank":0.31191623125197543,"base_score":2.0794415416798357,"endowment":2.0794415416798357,"self_citation_contribution":0.31191623125197543,"citation_network_contribution":0.0,"self_endowment_contribution":0.31191623125197543,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":7,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1676040,"name":"Biao Ma","orcid":null,"position":1,"is_corresponding":false},{"id":1139156,"name":"Jiali Li","orcid":"0000-0002-8198-1132","position":2,"is_corresponding":false},{"id":608842,"name":"Yaping Wang","orcid":"0000-0002-5382-3671","position":3,"is_corresponding":false},{"id":1676041,"name":"Tianyu Dou","orcid":null,"position":4,"is_corresponding":false},{"id":382821,"name":"Shujuan Xu","orcid":"0000-0001-5815-457X","position":5,"is_corresponding":false},{"id":1676042,"name":"Mingzhou Zhang","orcid":null,"position":6,"is_corresponding":false},{"id":1676039,"name":"Jungang Du","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Rapid Detection and Differentiation of Legionella pneumophila and Non-Legionella pneumophila Species by Using Recombinase Polymerase Amplification Combined With EuNPs-Based Lateral Flow Immunochromatography","abstract":"<jats:p><jats:italic>Legionella</jats:italic>, a waterborne pathogen, is the main cause of Legionnaires’ disease. Therefore, timely and accurate detection and differentiation of <jats:italic>Legionella pneumophila</jats:italic> and non<jats:italic>-Legionella pneumophila</jats:italic> species is crucial. In this study, we develop an easy and rapid recombinase polymerase amplification assay combined with EuNPs-based lateral flow immunochromatography (EuNPs-LFIC-RPA) to specifically distinguish <jats:italic>Legionella pneumophila</jats:italic> and non<jats:italic>-Legionella pneumophila</jats:italic>. We designed primers based on the <jats:italic>mip</jats:italic> gene of <jats:italic>Legionella pneumophila</jats:italic> and the 5S rRNA gene of non-<jats:italic>Legionella pneumophila.</jats:italic> The recombinase polymerase amplification reaction could go to completion in 10 min at 37°C, and the amplification products could be detected within 5 min with EuNPs-LFIC strips. Using a florescent test strip reader, the quantitative results were achieved by reading the colored signal intensities on the strips. The sensitivity was 1.6 × 10<jats:sup>1</jats:sup> CFU/ml, and a linear standard linear curve plotted from the test strip reader had a correlation coefficient for the determination of <jats:italic>Legionella pneumophila</jats:italic> (<jats:italic>R</jats:italic><jats:sup>2</jats:sup> = 0.9516). Completed concordance for the presence or absence of <jats:italic>Legionella pneumophila</jats:italic> by EuNPs-LFIC-RPA and qPCR was 97.32% (<jats:italic>κ</jats:italic> = 0.79, 95% CI), according to an analysis of practical water samples (<jats:italic>n</jats:italic> = 112). In short, this work shows the feasibility of EuNPs-LFIC-RPA for efficient and rapid monitoring of <jats:italic>Legionella pneumophila</jats:italic> and non-<jats:italic>Legionella pneumophila</jats:italic> in water samples.</jats:p>","is_dataset_classified":null,"base_score":2.0794415416798357,"endowment":2.0794415416798357,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"35198541","pmcid":"PMC8859533","openalex_id":"https://openalex.org/W4210942518","authors":[],"funders":[{"funder_name":"National Key Research and Development Program of China","grant_id":"2021YFF0602801","title":null}],"total_grants":1,"fwci":1.3459,"citation_percentile":0.76592162,"influential_citations":0,"citation_trend":[{"year":2023,"count":3},{"year":2024,"count":1},{"year":2025,"count":3}],"oa_status":"gold","license":"cc-by","oa_locations":[{"url":"https://www.frontiersin.org/articles/10.3389/fchem.2021.815189/pdf","host_type":"journal"},{"url":"https://www.frontiersin.org/articles/10.3389/fchem.2021.815189/pdf","host_type":"publisher"},{"url":"https://www.frontiersin.org/articles/10.3389/fchem.2021.815189/full","host_type":"publisher"},{"url":"https://doi.org/10.3389/fchem.2021.815189","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/35198541","host_type":"repository"},{"url":"https://doaj.org/article/00e31f776147471d8d40baada01c6c13","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/8859533","host_type":"repository"},{"url":"https://europepmc.org/articles/PMC8859533","host_type":"Europe_PMC"},{"url":"https://europepmc.org/articles/PMC8859533?pdf=render","host_type":"Europe_PMC"}],"fields_of_study":["Legionella and Acanthamoeba research","Biosensors and Analytical Detection","Vibrio bacteria research studies"],"mesh_terms":[],"keywords":["Legionella pneumophila","Recombinase Polymerase Amplification","Microbiology","Legionella","Polymerase chain reaction","Biology","Virology","Bacteria","Gene","Genetics","Simultaneous detection","Europium Nanoparticles","Fluorescent Immunochromatographic Assay"],"sdg_mappings":[{"sdg_number":0,"sdg_label":"Clean water and sanitation"}],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[{"name":"gen"},{"name":"refseq"},{"name":"doi"}],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-08T20:59:40.189241Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}