The Study Of Immunological Detecting Kits Of RSV And Their Applications

Rice stripe disease had broken out in Jiangsu province in recent years, which was caused by rice stripe virus (RSV). RSV was transmitted by small brown plant hopper (Laodelphax striatettus Fallen, SBPH), which could be transmitted in the permanent manner to offspring through ovary. The carrier rate of SBPH was one of the most significant factors for disease outbreak forecast. The present main methods for virus detecting were biological, immunological, and RT-PCR methods. Because of the advantage of speeding, handy, sensitive, accurate, lower expenditure, the immunological methods were the best in all. Based on monoclonal antibody of RSV, we started to study two detecting kits for RSV in SBPH and host plant, which based on the principal of indirect ELISA. The kits could be used for the study of RSV and SBPH, and also easily used for the agricultural technique extension. The application would provide first-hand data for the disease epidemic monitoring and forecast.We optimized the experimental condition of DIBA in laboratory from different dilution of samples, different concentration of antibody and different reaction time. The results suggested: at 37 ℃, the sample dilution was 100~200 times for SBPH and the infected leaves for 200-400 times. The dilution of monoclonal antibody was 1:20000-40000, reaction time for lh~1.5h, and Goat Anti-mouse IgG linked with Peroxides enzyme (second antibody) for 1:2000~2500, reaction time for 1.5h~2h. Then we searched for the 25 °C (out-lab) condition under 4°C, 25 °C, 37 °C, respectively reaction time for 1.5h and 3h in monoclonal antibody and Second disease. The best reaction time was confirmed, 3h was better than 1.5h. As for the results, we finished the kits and confirmed methods at 37°C and 25 °C (out-lab). And latterly, we detected expiring date of kits for 6 months. If the expiring date increased to 12 months, the dilution of Second disease should be increased for l:1000~1500.The kits were used to detect the females and their offspring to select high affinity population of SBPH in the fields from 13 counties in Jiangsu province. The high affinityindividuals had been obtained from all the districts, except for Guanyun, Gaoyou, Jianhu, Wujin counties. Among which, Hongze and Yandu counties had the high proportion in all, respectively for field population 15.29%, 17.39% and infected population 52%, 50%. A certain proportion of high affinity population was a crucial but not an only factor for the disease prevalence.The kits could be used to detect the carrier rate of SBPH from different areas, different fields of disease occurred, different generations, different life stages and different individual families in Jiangsu province. Compared with the results, we conclude that the carrier rate of SBPH was apparently different in different areas. The carrier rate of SBPH was lower than light diseased field. There were no certain relationships between the disease occurrence and the carrier rate of SBPH in a certain field. The variation of carrier rate looked like character "V" in over wintering generation, first and second generations, which indicated that RSV in SBPH decreased and then increased under field condition. With the growing of SBPH, the concentration of RSV in the body was raised. The lower concentration of RSV in body may make the baby nymph to live as easily as non-infected individuals. The trace of the carrier rate of SBPH in 66 families indicated that there are 30 families had 100% carrier rate in the 4th offspring, which owned 45%, and over 80% carrier rate of offspring owned 60%, But there were 40%~70% carrier rate of offspring own 20% and 20% families decreased to lower than 20% carrier rate. The carrier rate of offspring decreased differently in 8th generation, some to 0%. But 5 families also had 100% carrier rate. JY-116 family could maintain 100% in the 12th generation. After re-selected, the carrier rate of offspring could get back differently, some can completely recovery but some couldn't get back forever. As for the results, the interaction between SBPH an...