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利用环介导等温扩增技术与 FTA 技术相结合，快速、简单、直接检测感染根瘤的南方根结线虫。

Rapid, simple and direct detection of Meloidogyne hapla from infected root galls using loop-mediated isothermal amplification combined with FTA technology.

机构信息

State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China.

Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan 571101, China.

出版信息

Sci Rep. 2017 Apr 3;7:44853. doi: 10.1038/srep44853.

DOI:10.1038/srep44853
PMID:28368036
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5377304/
Abstract

The northern root-knot nematode (Meloidogyne hapla) is a damaging nematode that has caused serious economic losses worldwide. In the present study, a sensitive, simple and rapid method was developed for detection of M. hapla in infested plant roots by combining a Flinders Technology Associates (FTA) card with loop-mediated isothermal amplification (LAMP). The specific primers of LAMP were designed based on the distinction of internal transcribed spacer (ITS) sequences between M. hapla and other Meloidogyne spp. The LAMP assay can detect nematode genomic DNA at concentrations low to 1/200 000, which is 100 times more sensitive than conventional PCR. The LAMP was able to highly specifically distinguish M. hapla from other closely related nematode species. Furthermore, the advantages of the FTA-LAMP assay to detect M. hapla were demonstrated by assaying infected root galls that were artificially inoculated. In addition, M. hapla was successfully detected from six of forty-two field samples using FTA-LAMP technology. This study was the first to provide a simple diagnostic assay for M. hapla using the LAMP assay combined with FTA technology. In conclusion, the new FTA-LAMP assay has the potential for diagnosing infestation in the field and managing the pathogen M. hapla.

摘要

北方根结线虫（Meloidogyne hapla）是一种危害性很大的线虫，在全球范围内造成了严重的经济损失。本研究通过结合 Flinders 技术协会（FTA）卡和环介导等温扩增（LAMP），开发了一种用于检测侵染植物根系中 M. hapla 的敏感、简单、快速的方法。LAMP 的特异性引物是基于 M. hapla 和其他 Meloidogyne spp. 之间的内部转录间隔区（ITS）序列差异设计的。LAMP 检测可以检测到低至 1/200000 的线虫基因组 DNA，比常规 PCR 灵敏 100 倍。LAMP 能够高度特异性地从其他亲缘关系密切的线虫物种中区分出 M. hapla。此外，通过人工接种侵染根瘤的试验，证明了 FTA-LAMP 检测 M. hapla 的优势。此外，使用 FTA-LAMP 技术成功地从 42 个田间样本中的 6 个样本中检测到了 M. hapla。本研究首次利用 LAMP 联合 FTA 技术为 M. hapla 提供了一种简单的诊断检测方法。综上所述，新的 FTA-LAMP 检测方法有望用于田间侵染的诊断和管理 M. hapla 病原体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cea9/5377304/21d7d3a42c51/srep44853-f6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cea9/5377304/cc7fae4d1f46/srep44853-f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cea9/5377304/a8802755e5e6/srep44853-f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cea9/5377304/ae3ee157718b/srep44853-f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cea9/5377304/3dfef0336b9a/srep44853-f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cea9/5377304/40030c27a301/srep44853-f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cea9/5377304/21d7d3a42c51/srep44853-f6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cea9/5377304/cc7fae4d1f46/srep44853-f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cea9/5377304/a8802755e5e6/srep44853-f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cea9/5377304/ae3ee157718b/srep44853-f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cea9/5377304/3dfef0336b9a/srep44853-f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cea9/5377304/40030c27a301/srep44853-f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cea9/5377304/21d7d3a42c51/srep44853-f6.jpg

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