A set of oligonucleotides and probes for the detection and identification of viruses by the real-time PCR method

Abstract

The invention relates to molecular diagnostics and can be used in phytopathology, plant biotechnology, selection and plant breeding. It is a new set of synthetic oligonucleotides and probes for simultaneous detection of the genetic material of raspberry viruses such as raspberry ringspot virus (RpRSV), raspberry leaf spot virus (RLBV), raspberry bushy dwarf virus (RBDV) and raspberry leaf spot virus (RLMV) in field samples and in vitro plants. The real-time PCR method offers high sensitivity, specificity and speed of analysis, as well as the possibility of simultaneous detection of several pathogens and a low risk of contamination. The result of the invention is a set of oligonucleotides and probes for identification of viruses RRSV, RLSV, RBDV and RLBV in raspberry tissues. The design of oligonucleotides and probes was performed using multiple nucleotide sequences from international data banks to ensure interaction with all isolates of each virus. Approbation of the kit, using RNA isolated from raspberry leaves, showed high sensitivity and specificity of the method. The developed kit allows detection of RRSV, RLSV, RBDV and RLBV viruses in raspberry plants at all stages of vegetation, even without visible signs of viral infection.