Development of a hybridization capture HC-RT-PCR-ELISA method for reliable and sensitive detection of tomato ringspot virus

• Author(s): An, Derong; Chen, Hongyun; Huang, Wensheng; Zhao, Wenjun; Zhu, Shuifang;
• Abstract: Tomato ringspot virus is a very important quarantine virus (on the A1 list of China), which has not been reported from China yet. RT-PCR is a fast and sensitive method for ToRSV detection, but non-detectable and false positive result is often occurred. A novel hybridization capture RT-PCR-ELISA was developed based on the above mentioned RT-PCR. The main contributions to the development of the method are that a solid primer is successfully bound to PCR tube wall specially for aimed RNA capture in order to raise RNA purification and reduce time consumption. DIG labeled probe hybridization with solid PCR product was performed as well as electrophoresis of liquid product to reduce false positive result. The method was successfully used to detect the virus from the imported grape.
• Publication Date: Apr 2003
• Journal: Weishengwu Xuebao