Study On Technology Of Detecting Of Two Main Virus Of Potato With ELISA And RT-PCR

Potato is a kind of popular and important economic plant which grows in most of countries of the world. Potato is a staple food crop and widely grown in China. The diseases caused by viruses or viroids bring about substantial losses in agricultural production. There are more than 20 viruses and viroids which can infect potato. The proper diagnosis of causative viruses and viroids is the first requirement for the control of such diseases. Therefore, it is necessary of developing a rapid, sensitive and specific method for viruses detection. Potato virus A (PVA) and potato virus S (PVS) are top two viruses that result in reduction of potato yield in China.In the experiment, the detection methods of RT-PCR and ELISA were studied to detect PVA and PVS. Two methods were compared for isolation of total RNA from virus-infected and healthy potato leaves. Two pairs of DNA primers were designed and synthesized based on the nucleotide sequence of the coat protein (CP) gene of PVA and PVS respectively. The virus RNA was used for cDNA synthesis. Two specific PCR fragments about 678 bp and 642 bp were obtained by reverse transcription and polymerase chain reaction (RT-PCR) amplification, which was the expected length of PVA and PVS CP gene, while no fragment was obtained from the control in the same experiment. The results showed that a rapid and sensitive RT-PCR detection system of PVA and PVS was established, which could provide an effective way to detect two main virus.The technique of virus detection of PVS and PVA by DAS-ELISA were studied, and compared with RT-PCR method. The result showed that RT-PCR method was more sensitive and specific than ELISA method, however, ELISA method is more suitable for the detection of a great number of samples.