Rapid and accurate detection of Neofabraea perennans by droplet digital PCR
In order to realize high efficiency and high precision quarantine identification and epidemic monitoring of apple quarantine pathogen,the detection method of Neofabraea perennans by droplet digital PCR(ddPCR)was established.This method provides technical services and support for the prevention of pest from ports and the protection of China's ecological security.N.perennans was used as the research object,and the β-tubulin gene(TUB2)was identified by bioinformatics analysis as a target for specific primer and probe design.The specific detection system of N.perennans by ddPCR was established by optimizing the concentration of primers,probes and annealing temperature.The results showed that the detection limit of ddPCR was 0.16 copies/μL and the detection concentration of genomic DNA was 10.0 pg/μL,being consistent with the result of qPCR.The linear correlation coefficient in the range of quantitative detection was more than 0.999,and the RSD was less than 25%,indicating that the detection system had good repeatability and could be used for rapid and accurate detection of N.perennans in imported fruit samples.
droplet digital PCRapple bull's eye rotNeofabraea perennansrapid and accurate detection
万方数据
维普
