Method for PCR detection of pathogen of citrus greening disease

Abstract

The invention relates to a bioinstrumentation detection and identification method, in particular to a method for PCR detection of a pathogen of the citrus greening disease. The method for PCR detection of the pathogen of the citrus greening disease comprises the steps that DNA of a sample to be detected is extracted; a PCR amplification system is prepared, wherein the PCR amplification system comprises a primary PCR amplification system and a secondary PCR amplification system; a PCR amplification reaction is conducted; PCR amplification products are detected, wherein the final volume of the primary PCR amplification system and the final volume of the secondary PCR amplification system respectively ranges from 7microliters 10 microliters, wherein the final volume of a DNA template is 2.5 microliters, the final volume of 2*Taq PCR Master Mix is 2.5 microliters, the final volume of a primer is 0.25 microliter, and the rest is sterilization double distilled water. According to the method for PCR detection of the pathogen of the citrus greening disease, the final volume of the primary PCR amplification system and the final volume of the secondary PCR amplification system respectively ranges from 7microliters 10 microliters, the detection effect of a 20-microliter reaction system or a 25-microliter reaction system can be achieved, the economic cost of detection is greatly reduced, the amount of needed reagents is small, and the pollution to the environment is reduced.

Description

technical field [0001] The invention relates to a biological detection and identification method, in particular to a PCR detection method for the citrus huanglongbing pathogen. Background technique [0002] Citrus huanglongbing is one of the most serious diseases of citrus. It is listed as a key plant quarantine target at home and abroad. The pathogen is a fungus, which can be transmitted through grafting, seedling spread, and in the field through citrus psyllids. It not only turns the leaves yellow, but also reduces the yield and quality of citrus. In severe cases, the root system will rot and even destroy the entire orange orchard. It is an important control object in the management and protection of citrus. [0003] Conventional PCR detection technology only needs a pair of primers, and the accuracy is not high enough; due to the high price of various reagents and drugs required for the PCR detection reaction system of citrus Huanglongbing pathogen, many research institut...

AI Technical Summary

Problems solved by technology

[0003] Conventional PCR detection technology only needs a pair of primers, and the accuracy is not high enough; due to the high price of various reagents and medicines required for the PCR detection reaction system of citrus Huanglongbing pathogen, many scientific research institutes are unable to afford high detection costs due to limited funds. Although the citrus production departments in many places have the requirement to detect samples suspected of Huanglongbing, they often fail to complete the actual detection task indicators due to the lack of funds for detection, and the detection of complex geographical sources and massive samples is even more difficult to achieve. As a result, on the one hand, citrus greening disease has been spreading and developing year by year, but on the other hand, due to the high cost of sample testing, the practice and theory of citrus production are seriously out of touch.

At the same time, the reagents and drugs required for detection are toxic and will cause environmental pollution

At present, the total reaction system of the PCR detection technology for the Asian species of citrus greening disease is generally 20 μL or 25 μL, which requires a large amount of reagents and high resource consumption.

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