Study On The Rapid Detection Technique Of Pepper Mild Mottle Virus

In this study, pepper mild mottle virus(PMMoV) was purified, and the morphology was observedwith electron microscope, the moleculor weight was determined with SDS-PAGE. The PMMoV-specificpolyclonal antiserum was prepared for dot immunobinging assay (DIBA) and colloidal gold labellidimmunostrip (CGIS).The sensitivity and specificity of five methods including double antibodies sandwich enzyme-linkedimmuno sorbent assay (DAS-ELISA), DIBA, CGIS, RT-PCR and real-time fluorescent RT-PCR werecompared to detect PMMoV. The results as followings:(1)When separately from one infected seed with DAS-ELISA, CGIS, DIBA, RT-PCR and real-timefluorescent RT-PCR, the lowest detected dilution for the virus were 1:51200,1:51200,1:12800,1:100000,over 1:1000000. When infected leaves were detected, DAS-ELISA, CGIS and DIBA, the lowest detecteddilution were 1:3200.(2)The positive rate with PMMoV by DIBA were the different among 17 varieties from market inChina. Positive rate of 5 varieties were 0%, positive rate of 7 varieties were over 50%, and positive rate of3 varieties reached to 90% among detected 17 varieties.(3) Most of PMMoV was found in epidermis and testa, few in endosperm, not found in embryo byRT-PCR for PMMoV.(4) No PMMoV in seedlings from infected seeds and healthy seeds were detected out on seedling beds.Only 3 plants of 1280 seedlings from positive seeds were determined with PMMoV by DIBA aftertransplant, seed transmission rate was 0.23%. None of 1500 seedlings from infected 2 varieties and 1healthy variety were detected with PMMoV.