Real-time fluorescent PCR testing primer, probe and immobilized kit for citrus ulcer bacteria and testing process thereof

Abstract

This invention relates to oligonucleotide prime, mandarin orange ulcer germ real time fluorescence PCR detecting probe, solid phase reagent and its detecting method of mandarin orange ulcer germ detecting. It is a special quick detecting method direct towards mandarin orange ulcer disease, there is only 3 hours from sampling to detection, and detecting sensitivity is high, specificity is strong, stable and reliable. Misjudging in existing normal diagnosis method, missed diagnosis in electrical glass by asymmetry of germ and hard antibody preparation in single clone antibody and lack of specific reaction are conquered by this method, and false negative and false positive of normal PCR are avoided. Operation of solid phase kit is convenient, and its using is simple, and detecting sensitivity is high, specificity is strong, stable and reliable. So it is propitious to port detection of mandarin orange of import and export. Also it is propitious to allocation and transportation of mandarin orange inoculation, scion and other asymptomatic germ material, and early stage warning detection of non epidemic-stricken area orange ulcer disease.

Description

technical field [0001] The invention relates to a real-time fluorescent PCR solid-phase reagent kit and a quarantine inspection technology for the citrus canker pathogenic bacterium (Xanthomonas axonopodis pv.citri), which is a major pest. It belongs to the technical field of bioengineering. technical background [0002] Citrus canker is a major bacterial disease affecting the development of the global citrus industry, and it is also a key quarantine disease at home and abroad. Citrus canker fungus (Xanthomonas axonopodis pv.citri) is an important quarantine pest at home and abroad; it damages dozens of Rutaceae plants, including most commercial cultivars in Citrus parts (leaves, twigs, prickles, trunks, and fruit), while rarely directly killing the tree, can cause defoliation, fruit drop, tree weakening, and fruit canker lesions. [0003] The disease is mainly transmitted over long distances through the transportation of citrus materials such as seedlings, scions, and fru...

AI Technical Summary

Problems solved by technology

However, the disadvantages of the conventional method are that the detection cycle is long, the misjudgment rate is high for inexperienced people, and it cannot detect citrus asymptomatic materials, and it is difficult to meet the basic requirements of "fast, accurate and economical" plant quarantine.

In addition, the detection of citrus canker also includes phage test, enzyme-linked immunosorbent assay, dot immunosorbent assay, etc., which have problems such as complicated process, low sensitivity, high detection cost, and long cycle.