Detection Methods For Tomato Black Ring Virus And Cymbidium Mosaic Virus

Tomato black ring virus and Cymbidium mosaic virus are in risky pest list which is publicized by plant quarantine laws and regulations in China. At present, there are few reports for detection the two viruses on biology, electron microscope, serology, molecular biology detection as well as the molecular examination method sensitivity. Therefore, detection methods for the two Quarantine viruses were developed, the main results were as follows.1 TBRV was inoculated into some assay species, and in the end we selected a suit of host to distinguish TBRV.2 TBRV enzyme association immunity adsorption examination method was established; Immuno electron microscope method was established for the first time, by this way the virus particle was observed as sphere.3 RT-PCR and immuno capture RT-PCR (IC-PT-PCR) methods were established for the first time. The primers specific and MGB probe to the conserved sequence of TBRV coat protein gene were designed. The real time fluorescence PCR detection was firstly developed, with the Sensitivity of over 10-8μg/ml for Plasmind, it was enhanced 100 times compared to RT-PCR.4 Two pairs of primers specific to the conserved sequence of CymMV coat protein gene were designed, the RT-PCR, IC-PT-PCR as well as the sensitivity of RT-PCR were used to study the molecular detection method of CymMV. The results showed that CymMV could be confirmed by RT-PCR and IC-PT-PCR combined sequence determined. The sensitivity of CymMV by RT-PCR was determined using dilution of plasmid of CymMV and the minimum content of virus inspected was 2.72×10-7μg/ml , namely the examination smallest copy number was 428.