茄科作物重要病原物多目标现场快速检测方法的研究与应用OA

Strict seed quarantine is crucial for preventing the long-distance spread of seed-borne diseases in Solanaceae crops and ensuring agricultural biosafety.Crops like tomato and pepper hold high economic value and extensive indus-trial influence.However,devastating quarantine pathogens,including Clavibacter michiganensis subsp.michiganensis,Tomato leaf curl New Delhi virus,Tomato spotted wilt virus and Tomato brown rugose fruit virus,pose a severe threat to industry security.Current molecular detection techniques for the mixed bacterial and viral infections often suffer from complex procedures,reliance on specialized laboratories and personnel,and difficulties in achieving rapid,integrated on-site testing.To address the challenges,this study developed a fully automated,rapid on-site detection system based on microfluidic chip and real-time quantitative PCR technology.The system integrates modules for co-extraction of nucleic acids,coordinated multi-target amplification and visual interpretation of results.It achieves multiplex detection of the aforementioned four pathogens within a total process time of only 36 minutes.The constructed P1000F integrated de-tection system demonstrated limits of detection of 4.50×104 CFU/mL for the bacterial pathogen and 1.00×101 copies/μL for each of the three viral pathogens.Testing with actual samples confirmed its good applicability and reliability.Compared with current standard methods(EPPO standards and national standards),the detection sensitivity of this system was generally equivalent.In conclusion,the P1000F integrated detection method established in this study is efficient and accurate.It eliminates dependence on traditional laboratory environments and specialized technical personnel,providing reliable technical support for the rapid on-site screening and port quarantine of tomato seed-borne diseases.