Three Important Pathogensthe Establishment Of Nanozyme Strip Detection Technology

Lettuce mosaic virus(LMV)is a common pathogen that infects lettuce;Maize chlorotic mottle virus(MCMV)and A.citrulli are infectious crops pests;International exchanges and cooperation have caused the above three pathogens to be widely distributed worldwide.Based on the above,this study adopts the principle of double antibody sandwich method for the above three pathogens,through the combination of nano zyme coupling antibody and the antibody on the quality control line of nitrocellulose,and under the effect of diaminobenzidine(DAB),the color is developed,and the nanozyme test strip detection technology is established.The main results are as follows:(1)Establishment of the detection technology of the nanozyme strip of Lettuce mosaic virusDetection technology of nanozyme strip(DTNS)was established with Lettuce mosaic virus as the research object.The developed DTNS and enzyme linked immunosorbent assay(ELISA)are used to compare the sensitivity of purified LMV.The results showed that DTNS and ELISA can detect purified LMV down to 102 ng/mL and 103 ng/mL respectively.The former is 10 times more sensitive than the latter,which is indicating that the test strip has higher sensitivity,and at the same time the DTNS has the advantages of simpler operation and lower price than ELISA.Specificity experiments are performed by using the same genus and host as controls.The results show that only LMV was positive and 7 control viruses were negative,indicating that the established DTNS has good specificity.The established strips are used to test the samples,and the detection results are verified by ELISA and gene sequencing technique.The data shows that the three results are consistent.(2)Establishment of the detection technology of the nanozyme strip of Maize chlorotic mottle virusDetection technology of nanozyme strip(DTNS)was established with Maize chlorotic mottle virus as the research object..The established DTNS and the reverse transcription PCR(RT-PCR)technique are used to detect maize chlorotic mottle virus abrasives with a sensitivity of 10-7 and 10-5 respectively.Specific experiments are performed using the other four viruses as controls,there is no cross reactivity;Use the strip to test the sample,and verify the test results by RT-PCR technique and gene sequencing technique,the results show that the three are consistent.(3)Establishment of the detection technology of the nanozyme strip of A.citrulliDetection technology of nanozyme strip(DTNS)was established with A.citrulli virus as the research object.The nanozyme strip and colloidal gold test strip are used to detect the 10-fold gradient dilution of the bacterial liquid of A.citrulli.The result show that the minimum detection concentrations are 4.36 x 107 cfu/mL and 3.46 x 106 cfu/mL;Use 5 other bacteria as a control for specificity test,there is no cross reaction,so the specificity is good;Use nanozyme immunochromatographic strip and colloidal gold test strip to detect the leaf of the A.citrulli disease inoculated by acupuncture,the results are the same.The established nanozyme strip detection technology has the advantages of good specificity and high sensitivity,which can speed up the detection of pathogens and prevent the spread of pathogens in a large range in the field to a certain extent.This detection technology provides a new rapid detection method for pathogens.At the same time,the new material nanozyme has also been applied in the field of plant quarantine,which is of great significance for the protection of crops.