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松材线虫实时PCR检测技术(PDF)

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

Issue:
2007年04期
Page:
121-124
Column:
研究论文
publishdate:
2007-04-20

Article Info:/Info

Title:
Detection Technique of Bursaphelenchus xylophilus Using Real Time PCR
Article ID:
1000-2006(2007)04-0121-04
Author(s):
CHEN Feng-mao12 YE Jian-ren1* WU Xiao-qin1
1. College of Forest Resources and Environment Nanjing Forestry University, Nanjing 210037, China; 2. Anhui General Station of Forest Pest Management, Hefei 230031, China
Keywords:
Bursaphelenchus xylophilus Real-time PCR Specific primer Probe Detection
Classification number :
S763
DOI:
10.3969/j.jssn.1000-2006.2007.04.027
Document Code:
A
Abstract:
In this paper, a set of primers (F11/R11) and probe (TaqMan-11) specific for Bursaphelenchus xylophilus was designed to target the ribosomal DNA internal transcribed spacer (ITS) region, which composed the real-time polymerase chain reaction (PCR) assay. Then, B. xylophilus, B. mucronatus, B. hofmanni, A. macronucleatus and Seinura sp. separated from dead pine were detected a using this real-time PCR assay. The results showed that fluorescent signals were detected for all B. xylophilus isolates and there was no signal for B. mucronatus, B. hofmanni, Aphelenchoides macronucleatus, Seinura sp. and water (control). This indicated that the probe (TaqMan-11) and primers (F11/R11) were highly specific for B. xylophulus. Besides, the assay was very sensitive, detecting as little as 0.005 pg of B. xylophilus DNA in 10μL volume. The real-time PCR assay also successfully detected single pinewood nematodes, and it should be very useful for quarantine purposes.

References

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Last Update: 2013-05-20