การพัฒนาชุดตรวจสอบไวรัสใบหงิกของข้าวด้วย immunochromatographic assay

Abstract

Gold labeled IgG flow test (GLIFT) was developed as test kit for rice ragged stunt virus detection based on immunochromatographic assay. Accuracy, convenience and rapid detection are the novel properties of this technique. The GLIFT kit will be very useful for varietal screening in breeding program, forecasting and early warning of rice ragged stunt virus epidemic. The RRSV IgG was purified and tested by dot immunobinding assay (DIBA) to obtain the proper concentration for using in GLIFT kit. IgG was conjugated with colloidal gold and amount of 100-120 µl (6.6 µl/cm) of IgG conjugated gold were used for lining on 15 – 18 cm conjugated release pad (CRP). Control line and test line were made on nitrocellulose membrane using 40 µl (2.2 µl/ cm) of goat anti-rabbit (GAR) and IgG respectively. The appearance of color on both test line and control line indicated positive result. While no color was shown on the test line in case of negative result. GLIFT kit were tested for their efficacy to detect RRSV from infected rice leaves and viruliferous brown planthoppers. Satisfactory result was obtained with color gradually appeared on control line and test line within 5-10 minutes.