Method for rapidly detecting virus of potato tuber

Abstract

The invention discloses a method for rapidly detecting virus of a potato tuber. The method is characterized by comprising the following steps of: extracting the RNA of the potato tuber, performing reverse transcription-polymerase chain reaction (RT-PCR), detecting PCR products through a nucleic acid colloidal gold chromatography test strip, and the like. The method suitable for rapidly detecting the virus of the potato tuber is established by combining the RT-PCR technology with the nucleic acid colloidal gold chromatography technology on the premise of optimizing a method for extracting the RNA of the virus of the potato tuber. The method has the greatest characteristic that: an RNA extracting solution is developed, the RT-PCR detection can be directly performed after a potato sample to be detected is ground, an RNA extraction step in the conventional method is eliminated, and the problem that the RNA is difficult to extract due to high starch content of the potato tuber is solved; and moreover, the developed nucleic acid colloidal gold chromatography test strip can finish the detection of the PCR products within ten minutes, electrophoresis and the dyeing of ethidium bromide are not required, and the method is rapid, practical, safe and pollution-free.

Description

[0001] technical field [0002] The invention relates to a quick detection method of potato tuber virus, which mainly uses RT-PCR combined with nucleic acid immune colloidal gold chromatography test strip method to quickly detect potato tuber virus. [0003] Background technique [0004] Potato is the fourth largest food crop after wheat, corn and rice. It has wide distribution, strong adaptability, high yield, rich nutrition and multiple uses. The sown area and total output of potatoes in my country rank first in the world, but the per unit yield of potatoes in my country ranks 40th, which is in a relatively backward position. The reason is mainly due to the hazards of viruses (viruses). In recent years, the potato virus-free seed potato production technology represented by shoot tip tissue culture virus-free technology has been widely used, and has played a very significant role in the prevention and treatment of virus diseases. Rapid and sensitive virus detection is one...

AI Technical Summary

Problems solved by technology

However, due to the high starch content in potato tubers, it is difficult to extract RNA, so it is difficult to apply in the detection of potato tuber virus.

In addition, the routine detection method of PCR products is carried out by agarose gel electrophoresis. After electrophoresis, it needs to be stained with ethidium bromide and then observed under ultraviolet light. Ethidium bromide is moderately toxic and carcinogenic to humans, and it is easy to cause environmental pollution. , which also severely limits the generalization of this method

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