Detection kit for pine wood nematode and detection method therefor

Abstract

The invention discloses a case of rapid, sensitive, precise pine beam nematology detecting box and method, which consists of TaqMan-MGB probe and mating primer. The method adopts super-low temperature (-70 deg.c) freezing method to extract nematology DNA rapidly as form to proceed PCR augmentation, which judges the detecting result according to fluorescent curve. The invention is convenient to operate within 1.5 h to finish the whole detecting course, which is fit for animal quarantine, plant protection, packing company and forestry department.

Description

technical field [0001] The invention relates to the field of plant quarantine, in particular to a quarantine method for plant diseases and insect pests. Specifically, a method and kit for rapid detection of pine wood nematode using TaqMan-MGB probe real-time fluorescent PCR technology and its kit are provided, which are suitable for applications in plant quarantine, wood packaging production, plant protection and other departments. Background technique [0002] Pine wood nematode is the most dangerous forest disease in the world, and host plants can die within 40 days after being infected with pine xylophilus. Therefore, it is called the "cancer" of pine trees. It only takes 3-5 years from the onset to the destruction of the entire pine forest. The causative nematode of this disease is pine wood nematode (Bursaphelenchus xylophilus), which belongs to the order of the order, the superfamily, and the genus of the genus. At present, there is no good prevention and treatment ...

AI Technical Summary

Problems solved by technology

[0005] At present, the main quarantine method for pine wood nematodes at ports is morphological observation, but morphological observations have the following defects: First, pine wood nematodes intercepted at ports generally exist in the wooden packaging boards or backing boards of goods, and these materials are generally relatively dry , the number of nematodes is small, and most of them are larvae, which are often in a dormant state. Their stylets and esophagus have degenerated. It is difficult to directly identify them. Generally, they need to be cultured to adults before they can be identified.

However, there are still many defects in the detection method provided by this patent, mainly due to: (1) PCR method is too sensitive, easy to cross-contaminate, produces false positives, and lacks specificity; (2) PCR results need to be passed through gel electrophoresis To detect the results, the operation is cumbersome and time-consuming; the gel needs to be stained with EB, which is harmful to the human body and pollutes the environment

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