西北农业学报2026，Vol.35Issue(3)：580-588,9.DOI:10.7606/j.issn.1004-1389.2026.03.020
入侵害虫麦秆蝇(Elachiptera decipiens)LAMP检测方法的建立
Visual Identification of Invasive Pest Elachiptera decipiens Based on Ring-Mediated Isothermal Amplification Technique
摘要
Abstract
Elachiptera decipiens,an invasive insect pest recently identified in Xinjiang,damages wheat stems with its larvae and causes symptoms similar to those of other chloropid pests,making accurate differentiation difficult and complicating population monitoring.This study aimed to develop a rapid and visual detection method based on loop-mediated isothermal amplification(LAMP).The specific LAMP primers targeting the mitochondrial cytochrome oxidase Ⅰ gene(COⅠ)of E.decipiens were designed.The LAMP assay was then optimized,and the specificity and sensitivity of the primers were evaluated.A DNA toothpick extraction method combined with the optimized LAMP system was em-ployed for visual identification of samples.The results showed that the LAMP primers exhibited high specificity,and the optimized LAMP reaction system was as follows:2 μL of 10×Acculso Bca Buffer,1 μL of MgSO4(100 mmol/L),3.5 μL of dNTP Mix,1 μL of FIP/BIP(40 μmol/L),0.5 μL of F3/B3(10 μmol/L),3 μL of betaine(5 mol/L),1 μL of Bca DNA Polymerase,1 μL of Hydroxynaphthol Blue(250 μmol/L),9.5 μL of enzyme-free sterile water,and 1 μL of DNA template.The LAMP assay amplified positive samples within 50 min at constant-temperature 65℃,with a sensitive of very low levels of DNA(1×10-3 ng/μL).Combined with DNA toothpick extraction,the detection for samples could be completion within 1 h.Field sample testing demonstrates that the visual LAMP protocol is efficient,sensitive and simple to operate,satisfying the requirements for on-site detection during pest surveillance.关键词
入侵害虫/麦秆蝇/环介导等温扩增/线粒体CO Ⅰ基因/可视化鉴定Key words
Invasive pest/Elachiptera decipiens/Ring-mediated isothermal amplification/Mitochon-drial CO Ⅰ gene/Visual identification引用本文复制引用
张万龙,郭悦,戴琳,戴爱梅,张皓,胡祖庆..入侵害虫麦秆蝇(Elachiptera decipiens)LAMP检测方法的建立[J].西北农业学报,2026,35(3):580-588,9.基金项目
科技部对发展中国家援助项目(KY202002018) （KY202002018）
国家现代农业产业技术体系项目(CARS-03-37) （CARS-03-37）
新疆维吾尔自治区自然科学基金(2022D01A06).Assistance Programs for Developing Countries by Ministry of Science and Technolo-gy of China(No.KY202002018) （2022D01A06）
National Modern Agricultural Industry Technology System(No.CARS-03-37) （No.CARS-03-37）
Natural Science Foundation of Xinjiang Uygur Autonomous Region(No.2022D01A06). （No.2022D01A06）
