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

Potato is a staple food crop and widely grown in China.Virus is a main factor that affects the potato production.Potato virus X(PVX), potato virus Y(PVY), and potato leaf roll virus(PLRV) are top three viruses that result in decreased yield of potato in china.Therefore,the rapid methods of ELISA and RT-PCR were studied to detect PVX, PVY, and PLRV. The potato leaves infected with PVX, PVY, and PLRV were used as dornors,the techniques of virus detecting them by DAS-ELISA were studied and an integrated detecting program were established. In the experiment,three pairs of DNA primers were designed and synthesized based on the nucleotide sequence of the coat protein (CP) gene of PVX, PVY, and PLRV respectively. The virus RNA directly extracted from virus-infected potato leaves was used for cDNA synthesis. Three specific PCR fragments about 535bp,680b and 627 bp were obtained by reversetranscription and polymerase chain reaction (RT-PCR) amplification,which was the expected length of PVX,PVY and PLRV 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 PVX,PVY and PLRV was established,which could provide an effective way to detect three main virus. Two simple, rapid, sensitive, reliable detection techniques of three virus are reported in the paper: they are DAS-ELISA and RT-PCR. The virus RNA without purification could be detected with ELISA technique,which is simpler and rapider than RT-PCR technique, and is more suitable for the detection of a great number samples. The sensitivity of ELISA is as same as that of RT-PCR,but RT-PCR technique is more specific.