biostudies-literatureUnknown13Yang X<i>Brucella abortus</i> (<i>B. abortus</i>) as an important infectious agent of bovine brucellosis cannot be ignored, especially in countries/regions dominated by animal husbandry. Thus, the development of an ultrasensitive and highly specific identification technique is an ideal strategy to control the transmission of bovine brucellosis. In this report, a novel detection protocol, which utilizes multiple cross displacement amplification (MCDA) combined with a gold nanoparticles-based lateral flow biosensor (AuNPs-LFB) targeting the <i>BruAb2_0168</i> gene was successfully devised and established for the identification of <i>B. abortus</i> (termed <i>B. abortus</i>-MCDA-LFB). Ten specific primers containing engineered C1-FAM (carboxyfluorescein) and D1-biotin primers were designed according to the MCDA reaction mechanism. These genomic DNA extracted from various bacterial strains and whole blood samples were used to optimize and evaluate the <i>B. abortus</i>-MCDA-LFB assay. As a result, the optimal reaction conditions for the <i>B. abortus</i>-MCDA-LFB assay were 66°C for 40 min. The limit of detection of the <i>B. abortus</i>-MCDA-LFB was 10 fg/μl (~3 copies/μl) for genomic DNA extracted from pure cultures of <i>B. abortus</i> isolate. Meanwhile, the <i>B. abortus</i>-MCDA-LFB assay accurately identified all tested <i>B. abortus</i> strains, and there was no cross-reaction with non-<i>B. abortus</i> pathogens. Moreover, the detection workflow of the <i>B. abortus</i>-MCDA-LFB assay for whole blood samples can be completed within 70 min, and the cost of a single test is approximately 5.0 USD. Taken together, the <i>B. abortus</i>-MCDA-LFB assay is a visual, fast, ultrasensitive, low-cost, easy-to-operate, and highly specific detection method, which can be used as a rapid identification tool for <i>B. abortus</i> infections.Frontiers in microbiology1071928https://www.ebi.ac.uk/biostudies/studies/S-EPMC9744775biostudies-literatureRapid, ultrasensitive, and highly specific identification of <i>Brucella abortus</i> utilizing multiple cross displacement amplification combined with a gold nanoparticles-based lateral flow biosensor.PMC9744775Yang XLiu YLi SHuang JWei XZeng XYing XWang YTan QfalseRapid, ultrasensitive, and highly specific identification of <i>Brucella abortus</i> utilizing multiple cross displacement amplification combined with a gold nanoparticles-based lateral flow biosensor.<i>Brucella abortus</i> (<i>B. abortus</i>) as an important infectious agent of bovine brucellosis cannot be ignored, especially in countries/regions dominated by animal husbandry. Thus, the development of an ultrasensitive and highly specific identification technique is an ideal strategy to control the transmission of bovine brucellosis. In this report, a novel detection protocol, which utilizes multiple cross displacement amplification (MCDA) combined with a gold nanoparticles-based lateral flow biosensor (AuNPs-LFB) targeting the <i>BruAb2_0168</i> gene was successfully devised and established for the identification of <i>B. abortus</i> (termed <i>B. abortus</i>-MCDA-LFB). Ten specific primers containing engineered C1-FAM (carboxyfluorescein) and D1-biotin primers were designed according to the MCDA reaction mechanism. These genomic DNA extracted from various bacterial strains and whole blood samples were used to optimize and evaluate the <i>B. abortus</i>-MCDA-LFB assay. As a result, the optimal reaction conditions for the <i>B. abortus</i>-MCDA-LFB assay were 66°C for 40 min. The limit of detection of the <i>B. abortus</i>-MCDA-LFB was 10 fg/μl (~3 copies/μl) for genomic DNA extracted from pure cultures of <i>B. abortus</i> isolate. Meanwhile, the <i>B. abortus</i>-MCDA-LFB assay accurately identified all tested <i>B. abortus</i> strains, and there was no cross-reaction with non-<i>B. abortus</i> pathogens. Moreover, the detection workflow of the <i>B. abortus</i>-MCDA-LFB assay for whole blood samples can be completed within 70 min, and the cost of a single test is approximately 5.0 USD. Taken together, the <i>B. abortus</i>-MCDA-LFB assay is a visual, fast, ultrasensitive, low-cost, easy-to-operate, and highly specific detection method, which can be used as a rapid identification tool for <i>B. abortus</i> infections.2022-01-01T00:00:00Z20222025-04-04T12:01:14.868Z2025-02-19T02:59:49.99ZS-EPMC97447753652383010.3389/fmicb.2022.1071928