PCR DETECTING METHOD BARLEY YELLOW MOSAIC VIRUS FROM SOIL USING NESTED PCR

摘要

The present invention relates to a method of detecting the barley yellow mosaic virus (BaYMV) from soil by using nested PCR. In particular, the method comprises: a first step of isolating RNA from a soil sample obtained from the soil near the barley being cultivated; a second step of synthesizing cDNA by using a reverse transcriptase while using the RNA isolated in the first step as a template; a third step of performing primary PCR with a primer combination of SEQ ID NO: 1 and SEQ ID NO: 2 while using the cDNA synthesized in the second step as a template; a fourth step of performing secondary PCR with a primer combination of SEQ ID NO: 3 and SEQ ID NO: 4 while using the primary PCR product as a template; and a fifth step of identifying BaYMV genes from the secondary PCR product. The method of the present invention is an effective detection technique with significantly higher specificity and sensitivity compared to a conventional method, and thus may be used to diagnose microorganisms parasitic on other microorganisms or difficult-to-detect pathogens with extremely low habitat density. Furthermore, the method of the present invention may be advantageously used as a tool to diagnose with precision the presence of soil-borne pathogens in the field of quarantine and inspection.