Open AccessDetection of the Pinewood Nematode, Bursaphelenchus xylophilus, Using a Real-Time Polymerase Chain Reaction Assay
Author(s) -
Aixin Cao,
X Z Liu,
Shuifang Zhu,
Bingyue Lu
Publication year - 2005
Publication title -
phytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.264
H-Index - 131
eISSN - 1943-7684
pISSN - 0031-949X
DOI - 10.1094/phyto-95-0566
Subject(s) - bursaphelenchus xylophilus , xylophilus , biology , polymerase chain reaction , primer (cosmetics) , internal transcribed spacer , real time polymerase chain reaction , nematode , ribosomal rna , ribosomal dna , microbiology and biotechnology , botany , gene , genetics , ecology , phylogenetics , chemistry , organic chemistry
The pinewood nematode, Bursaphelenchus xylophilus, has caused significant damage to pine plantations both in East Asia and North America and is an important quarantine organism. A real-time polymerase chain reaction (PCR) assay was developed to detect B. xylophilus. A set of primers and probe specific for B. xylophilus was designed to target the ribosomal DNA internal transcribed spacer region. Optimal primer concentration, Mg 2+ concentration, and extension temperature were 400 nM, 3.0 mM, and 60°C, respectively. The assay was highly specific and sensitive, detecting as little as 0.01 ng of B. xylophilus DNA. The real-time PCR assay also successfully detected B. xylophilus in field samples, and it should be very useful for quarantine purposes.