Ahuja Amit, Joshi Vijay, Singh Gagandeep, Kundu Artha, Bhat Chaitra G, Kumar Satya, Rao Uma, Somvanshi Vishal Singh
Division of Nematology, ICAR-Indian Agricultural Research Institute, New Delhi, 110012 India.
Department of Plant Pathology, College of Agriculture, G. B. Pant University of Agriculture and Technology, Udham Singh Nagar, Pantnagar, Uttarakhand 263145 India.
3 Biotech. 2021 Jun;11(6):294. doi: 10.1007/s13205-021-02830-8. Epub 2021 May 24.
Cyst nematodes of the species and are devastating parasites of the potato crop. Early detection of cyst nematodes in the field is critical for adopting an appropriate management strategy. A specific and sensitive loop-mediated isothermal amplification (LAMP) assay using four oligonucleotide primers has been developed to amplify the internal transcribed spacer region (ITS) of ribosomal DNA of potato cyst nematode The PCN-LAMP reaction could be completed within 75 min at 68 °C followed by termination at 85 °C for 7 min. The primers exhibited specificity for and did not detect any other tested genera of plant parasitic or entomopathogenic nematodes. LAMP reaction was highly sensitive, suitable for crude genomic DNA and could successfully detect DNA up to femtogram quantity. This assay is rapid, cost effective and requires minimal instrumentation. It will facilitate the detection of at field and point-of-care labs and help in the interception of infested plant material/soil samples at quarantine stations independent of a professional nematologist.
The online version contains supplementary material available at 10.1007/s13205-021-02830-8.
属和 属的孢囊线虫是马铃薯作物的毁灭性寄生虫。在田间早期检测孢囊线虫对于采用适当的管理策略至关重要。已开发出一种使用四种寡核苷酸引物的特异性和灵敏的环介导等温扩增（LAMP）检测方法，以扩增马铃薯孢囊线虫核糖体DNA的内部转录间隔区（ITS）。马铃薯孢囊线虫LAMP反应可在68℃下75分钟内完成，然后在85℃下终止7分钟。这些引物对 属和 属具有特异性，未检测到任何其他测试的植物寄生或昆虫病原线虫属。LAMP反应高度灵敏，适用于粗基因组DNA，能够成功检测至飞克量的 属DNA。该检测方法快速、经济高效且所需仪器最少。它将有助于在田间和即时检测实验室检测 属，并有助于在检疫站拦截受侵染的植物材料/土壤样本，而无需专业线虫学家。
在线版本包含可在10.1007/s13205-021-02830-8获取的补充材料。
