Journal
PHYTOPATHOLOGY
Volume 95, Issue 5, Pages 566-571Publisher
AMER PHYTOPATHOLOGICAL SOC
DOI: 10.1094/PHYTO-95-0566
Keywords
cycle threshold; normalized fluorescence
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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, Mg2(+) concentration, and extension temperature were 400 nM, 3.0 mM, and 60 degrees 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. xylophilits in field samples, and it should be very useful for quarantine purposes.
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