Colloidal gold immunization test strip and detection method for detecting plum pox virus

Abstract

The invention discloses a colloidal gold immunization test strip and a detection method for detecting plum pox viruses. The colloidal gold immunization test strip is formed by the steps of: A. extracting RNA (Ribonucleic Acid) of plum pox viruses from materials infected by the plum pox viruses; B. cloning coat protein genes of the plum pox viruses by an RT-PCR (Reverse Transcription-Polymerase Chain Reaction) method, connecting the coat protein genes on a prokaryotic expression vector pET29(a) to obtain pET29a-PPV (porcine parvovirus) recombinant vectors, orderly measuring and finally verifying the recombinant vectors with correct reading frames, carrying out the inducible expression with the coat protein genes of the plum pox viruses, and transforming escherichia coli BL21 (DE3) with the pET29a-PPV recombinant vectors; C. recovering specific expression protein by an affinity column method, and making the rabbits immune to obtain plum pox virus antiserum; and D. adopting the above antiserum to prepare the colloidal gold immunization test strip used for detecting the plum pox viruses by the prepared antiserum. The invention aims to provide the convenient colloidal gold immunization test strip for rapidly detecting the plum pox viruses and the method for detecting plum pox viruses by adopting the test strip.

Description

technical field [0001] The invention relates to a colloidal gold immune test strip for detecting plum pox virus, and also relates to a method for detecting plum pox virus by using the colloidal gold immune test strip. Background technique [0002] Plumpox virus (PPV) is the latest quarantine pathogen promulgated by my country, belonging to the Potyvirus genus (Potyvirus) in the family Potyviridae (Potyviridae), and it is one of the most dangerous viruses for stone fruit trees (Dunez , Sutic, 1988; Nemeth, 1994), its natural hosts are mainly woody plants of the genus Prunus, such as apricot (PrunusarmeniacaL.), peach (P.persicaL.) and plums (P.domesticaL. and P. salicinaLindl.) and sweet cherry (P.aviumL.) and sour cherry (P.cerasusL.) and so on. Once the fruit tree is infected by it, the leaves will be distorted and chlorotic, the veins will be yellowed, especially the fruit will be deformed and become smaller, and mottled will appear, resulting in a decline in quality, and a...

AI Technical Summary

Problems solved by technology

Although enzyme-linked immunosorbent assay (ELISA) is still the main method for detection of fruit tree virus diseases at ports, ELISA requires specialized equipment and professionals to perform detection and judgment. The operation is cumbersome and takes a long time , there is an urgent need for a simpler, faster and more sensitive detection method to be more suitable for the requirements of quick inspection and release at ports

[0004] At present, the detection methods of plant viruses in foreign countries mainly include the following five kinds: one is the traditional biological method, which is characterized by long cycle, low accuracy, small number of detected viruses, and high labor intensity; the other is the electron microscope method, but It can only observe the shape of virions, but cannot identify the virus species, and the operation is complicated and the price is very expensive; the third is enzyme-linked immunosorbent assay, which is the most widely used virus detection method at present, but its detection time is long and the operation steps are many , and requires special instruments and the purchase of expensive specific antiserum; Fourth, although the PCR method is very sensitive and fast, it requires special instruments and professionals, and the detection throughput is limited; Fifth, gene chip detection technology, which is currently The most promising and potential detection method, but the stability and repeatability of the detection results need to be further improved, and the price of the instrument is also very expensive, the operation is complicated, and the detection time is long

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