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可视化环介导等温扩增技术检测 Phytophthora tentaculata(PDF)

《南京林业大学学报(自然科学版)》[ISSN:1000-2006/CN:32-1161/S]

Issue:
2015年06期
Page:
1-6
Column:
专题报道
publishdate:
2015-11-30

Article Info:/Info

Title:
Rapid diagnostic methods for Phytophthora tentaculata using LAMP
Article ID:
1000-2006(2015)06-0001-06
Author(s):
DAI Tingting12 KANG Zhenye12 WU Xiaoqin12
1. Co-Innovation Center for the Sustainable Forestry in Southern China, Nanjing 210037, China;
2. Jiangsu Key Laboratory for Prevention and Control of Invasive Species, College of Forestry, Nanjing Forestry University, Nanjing 210037,China
Keywords:
Phytophthora tentaculata loop-mediated isothermal amplification(LAMP) PtYpt1 hydroxynaphthol blue
Classification number :
S763
DOI:
10.3969/j.issn.1000-2006.2015.06.001
Document Code:
A
Abstract:
In order to identify the disease caused by Phytophthora tentaculata, and keep the spread of the disease under control and reduce the economic losses quickly and accurately, loop-mediated isothermal amplification(LAMP)assay targeting the PtYpt1 element was used to detect Phytophthora tentaculata. The PtYpt1-LAMP assay amplified the target element in less than 80 min at 64 ℃ efficiently and was evaluated specificitily and sensitivitily. The specificity was evaluated against Phytophthora tentaculata, Phytophthora spp., Pythium spp., and true fungi isolates. The P. tentaculata DNA products were visualised as a ladder-like banding pattern by 2% gel electrophoresis. A positive colour(sky blue)was only observed in the presence of P. tentaculata by addition of hydroxynaphthol blue prior to amplification, whereas none of other isolates showed a colour change. The detection limit of the PtYpt1-specific LAMP assay for P. tentaculata was 1 ng/μL of genomic DNA per reaction.

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Last Update: 2015-11-30