RT-PCR detection method of ordinary rice black streaked dwarf virus (RBSDV)

Abstract

The invention relates to an RT-PCR detection method of an ordinary rice black streaked dwarf virus (RBSDV). A pair of used primers are obtained by the method that the first section of RNA of an RBSDV is specific, and nucleotide sequences of the first section of RNA of the RBSDV and the first section of RNA of similar viruses are researched and downloaded from NCBI by utilizing the conservative property of the evolution of viral nucleotide sequences; and the sequences are compared by using MEGA4.0 software to find out a primer candidate area which is reported by different researchers and shared by the RBSDV nucleotide sequences and is different from other viral sequences from a mas file. Proved by BLAST results, the sequences which can 100 percent cover the primer 1 and the primer 2 and 100 percent same as the primer 1 and the primer 2 are all the RBSDV nucleotide sequences. RBSDV and SRBSDV viruses can be exactly separated by using the pair of primers for carrying out RT-PCR detections on the RBSDV and SRBSDV viruses.

Description

Technical field: [0001] The invention relates to a detection method of common rice virus, in particular to a RT-PCR (reverse transcription polymerase chain reaction) detection method of common rice black streaked dwarf virus (RBSDV). Background technique: [0002] Ordinary rice black-streaked dwarf virus disease is a serious viral disease in Jiangsu and Zhejiang, causing serious losses every year. In 2008, South China Agricultural University and Zhejiang Academy of Agricultural Sciences respectively reported that a new viral disease occurred in Guangdong and Hainan provinces. South China Agricultural University tentatively named the virus as southern rice blackstreaked dwarf virus. , SRBSDV), Zhejiang Academy of Agricultural Sciences tentatively named rice black streaked dwarf virus 2 (rice black streaked dwarf virus 2, RBSDV 2). In 2009, dwarf disease similar to these two viral diseases occurred in a large area in Hunan Province. According to preliminary statistics, the af...

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Problems solved by technology

[0003] Regarding the detection technology of RBSDV, Jiangsu Academy of Agricultural Sciences and other research units have previously designed a pair of primers for RT-PCR detection of the disease based on the nucleotide sequence of the first RNA fragment of the virus, but the inventor found in the actual detection work This is not good for the detection of primers. First, the electrophoretic bands are not very clear when detecting RBSDV-positive samples; second, the specificity is not good. The expected band of about 200bp was expanded in the Zhejiang Puhei sample, while the one in Hunan has been proved to be The SRBSDV specimen also extended a band of about 1500bp, see figure 1

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