DETECTION OF VIROID OF POTATO STREPTOMYCES SCABIES AND PROBE FOR ITS DETECTION

摘要

PURPOSE: To detect the potato Streptomyces scabies viroid(PSTVd) with high sensitivity for preventing potatoes from being infected with the viroids by using a plurality of synthetic oligonucleotides which are complementary to the PSTVd RNA as probes. ;CONSTITUTION: Synthetic oligonucleotides having bases sequences represented by formulas I to V which are complementary to the RNA of PSTVd are used in a mixture as probes. Raw leaves of tomato or the like infected with PSTVd are ground to extract the nucleic acid soluble in 2M lithium chloride, a reverse transcriptase is allowed to act on to synthesize cDNA, which is amplified by PCR. The amplified cDNA is immobilized to the microplate and a mixture of biotin-labeled synthetic oligonucleotides of formulas I to V is hybridized to the cDNA. Then, avidinated alkali phosphatase is allowed to react with the hybridomas and a color-developing substrate is allowed to act on the reaction mixture to determine PSTVd colorimetrically using a spectrophotometer.;COPYRIGHT: (C)1996,JPO