Pinewood nematoda detection reagent-box and detection method

Abstract

Two pairs of primer are designed: Pl / P3 is an idiocratic primer for pine wood nematode; P2 / P3 an idiocratic primer for quasi pine wood nematode. The kit includes 1.25ml solution I,1ml solution II, 100U tag enzyme, and 50ul for each contrapositive DNA of wireworm. The solution I is as following constitutes: Worm Lysis Buffer, 500mM KCl, 100mMTris-ClpH 8.0, 15mM MgCl2, 10mMDTT, 4.5% Tween20, 0.1%gelatin, 0.2ug / ul protease K. The solution II is as following constitutes: 2xPCR reaction Buffer, 4.0mM MgCl2, 0.2mM dNTP, 0.8 mu MP1, P2, P3. The invented kit possesses high specificity, sensitiveness and stability, providing fast, sensitive and specific technical method.

Description

(1) Technical field [0001] The invention relates to a pine wood nematode (Bursaphelenchus xylophilus) detection kit and a detection method thereof, belonging to the field of biotechnology. It is specially used for the high-sensitivity and rapid detection of pine wood nematodes carried by customs inbound and outbound timber and wooden packaging. At the same time, it can be used for the detection and diagnosis of pine wood nematode disease. (2) Background technology [0002] Pine wood nematode disease, also known as pine wilt disease, is a devastating disease in forestry, known as forest cancer. Infected pine trees are infected by it, and when the environmental conditions are suitable, the whole plant dies in only about one month. The disease was first discovered in Nagasaki, Kyushu, Japan in 1905, and then reported in the United States, Canada, Mexico, South Korea, Portugal and other places. The pathogenic nematode of the disease is pine wood nematode [Bursaphelenchus xylo...

Images