The Molecular Identification And DAS-ELISA Detection Of Three Potato Viruses In Guizhou

Potato is a staple food crop and widely grown in the world. Virus is a main factor that affects great loss of the potato production and is the main reason that leads to potato degeneration. 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 DAS-ELISA was studied to test 202 samples of potato leaves gathered from potato planting areas in Guizhou province PVX, PVY and PLRV.In the experiment, three pairs of DNA specific primers were designed and synthesized based on sequence of the coat protein (CP) gene of PVX, PVY, and PLRV respectively. The virus RNA directly extracted or immunocapture from PVY infected potato leaves was used for cDNA synthesis. The specific PCR fragments about 804bp was obtained by reverse transcription and polymerase chain reaction(RT-PCR) amplification, which was the expected length of PVY CP Gene. Then using IC-RT-PCR method ,the specific PCR fragments about 714bp and 627bp were obtained by reverse transcription and polymerase chain reaction(RT-PCR) amplification, which were also the expected length of PVX and PLRV CP Gene. the phylogenic trees of PVX,PVY,PLRV were established Based on CP nucleotide acid. Accession numbers of these sequences were gotten which were submitted to National Center for Biotechnology Information(NCBI) are EF063709,EF063710,EF063711 and EF063712.Tree simple, rapid, sensitive, reliable detection techniques of three virus arereported in the paper: they are DAS-ELISA,RT-PCR and IC-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 numbersamples. The sensitivity of ELISA is as same as that of RT-PCR. But RT-PCR and IC-RT-PCR technique is more specific. IC-RT-PCR without the step of extracting RNA is more simple than RT-PCR. The sensitivity of IC-RT-PCR is as same as that of RT-PCR.