| 10 | 0 | 14 |
| 下载次数 | 被引频次 | 阅读次数 |
为建立快速鉴别樱桃绕实蝇的现场检测方法，本研究利用樱桃绕实蝇线粒体细胞色素COI基因序列设计特异的环介导等温扩增引物，建立了基于LAMP的微流控芯片检测方法。结果表明，LAMP微流控芯片方法特异性好，与近似种样本无交叉反应；对樱桃绕实蝇的检测灵敏度可达1 pg/μL，可在30 min内完成，测试樱桃绕实蝇基因组DNA重复性Ct值变异系数（CV）均在2%以下，稳定性好。本方法的建立为樱桃绕实蝇的现场快速检测提供了有力工具。
Abstract：To establish a rapid on-site detection method for R. cerasi,this study designed specific loop-mediated isothermal amplification(LAMP) primers using the mitochondrial cytochrome COI gene sequence of R. cerasi and developed a microfluidic chip detection method based on LAMP. The results showed that the LAMP microfluidic chip method has good specificity and does not cross-react with closely related species. Its sensitivity in detecting R. cerasi can reach 1 pg/μL and can achieve within 30 minutes. The coefficient of variation(CV) of repeated Ct values for cherry fruit fly genomic DNA tests was below 2%,indicating good stability. The establishment of this method provides a powerful tool for rapid on-site detection of R. cerasi.
[1]DANIEL C,GRUNDER J. Integrated management of european cherry fruit fly Rhagoletis cerasi(L.):Situation in Switzerland and Europe. Insects,2012,3(4):956-988.
[2]LAWRENCE B. First record of the European cherry fruit fly,Rhagoletis cerasi(Linnaeus)(Diptera:Tephritidae),in North America. Insecta Mundi,2018,0622:1-4.
[3]WAKIE T T,YEE W L,NEVEN L G. Assessing the risk of establishment of Rhagoletis cerasi(Diptera:Tephritidae)in the United States and globally. J Econ Entomol,2018,111(3):1275-1284.
[4]SVETOMIR S,PANTELIJA P,DRAGO M. Rhagoletis cerasi Loew(Diptera:Tephritidae)biological characteristics,harmfulness,and control. Pesticidi i Fitomedicina,2012,27(4):269-281.
[5]秦誉嘉，蓝帅，卢国彩，等.气候变化条件下樱桃绕实蝇在中国的潜在地理分布预测.植物保护学报，2019,46(1):63-70.
[6]NOTOMI T,OKAYAMA H,MASUBUCHI H,et al. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res,2000,28(12):E63.
[7]王华，余海龙，任毓忠.苹果黑星菌LAMP快速检测方法的建立.中国农学通报，2025,41(3):139-144.
[8]张万龙，郭悦，戴琳，等.入侵害虫麦秆蝇（Elachiptera decipiens）LAMP检测方法的建立[J/OL].西北农业学报，[2026-01-15].https：//link.cnki.net/urlid/61.1220.S.20250427.1535.002.
[9]刘玮琦，赵畅，姜帆，等.基于LAMP技术的欧洲樱桃绕实蝇检测方法的建立.植物检疫，2024,38(6):23-30.
[10]俞禄珍，李帅，林宇，等.草莓滑刃线虫LAMP检测方法的建立.植物检疫，2025,39(1):35-38.
[11]姜宁，白洁，李平，等.基于环介导等温扩增微流控芯片技术快速检测蚊媒携带登革病毒方法的建立.中国寄生虫学与寄生虫病杂志，2024,42(2):234-241.
[12]石艳萍，邓飞，周丽媛，等.微流控芯片环介导等温扩增技术检测3种猪圆环病毒.中国动物检疫，2024,41(2):58-64.
[13]尚玉婷，叶青华，蔡淑珍，等.基于环介导等温扩增技术的微流控芯片在病原菌检测方面的研究进展.微生物学报，2021,61(10):2999-3009.
[14]KEPENEKCİİ,HAZIR S,ÖZDEM A. Evaluation of native entomopathogenic nematodes for the control of the European cherry fruit fly Rhagoletis cerasi L.(Diptera:Tephritidae)larvae in soil. Turkish Journal of Agriculture and Forestry,2015,39:74-79.
[15]刘玮琦，姜帆，李兰，等. Taq Man探针实时荧光定量PCR快速鉴定欧洲樱桃绕实蝇.植物检疫，2022,36(3):39-45.
[16]NORMAN B B,DANIEL G,LISA A L,et al. Using the r DNA internal transcribed spacer 1 to identify the invasive pest Rhagoletis cerasi(Diptera:Tephritidae)in North America.Journal of Economic Entomology,2021,114(1):360-370.
基本信息:
DOI：10.19662/j.cnki.issn1005-2755.2026.02.012
中图分类号:S41-30
引用信息:
[1]刘玮琦,赵畅,李兰,等.基于环介导等温扩增的微流控芯片技术检测樱桃绕实蝇[J].植物检疫,2026,40(02):71-77.DOI:10.19662/j.cnki.issn1005-2755.2026.02.012.
基金信息:
“天山英才”培养计划青年拔尖人才项目（2023TSYCJC0020）; 海关总署科研项目（2021HK171;2022HK014;2024HK231）; 陕西省重点研发计划项目（2024GX-YBXM-547）