RT-PCR detection method of south rice black-streaked dwarf virus (SRBSDV)

Abstract

The invention relates to an RT-PCR detection method of a south rice black-streaked dwarf virus (SRBSDV). In the detection method, SRBSDV specific primers are used for carrying out RT-PCR amplification on a sample to be detected and then carrying out electrophoretic identification. The specific primers are obtained by the method that by utilizing the conservatism of the evolution of viral nucleotide sequences, the tenth section of RNA of SRBSDV and nucleotide sequences of the tenth section of RNA of similar viruses are searched and downloaded from NCBI / GenBank, the sequences are compared by utilizing MEGA4.0 software, and then a pair of primers which are shared by all the SRBSDV nucleotide sequences reported by different researchers and are different from other viral sequences are found out from a file with an extension name of mas. Proved by a BLAST result, sequences which 100 percent cover the primers and 100 percent same as the primers are all the SRBSDV nucleotide sequences. The primers can be used for accurately separating SRBSDV diseased plants from ordinary RBSDV diseased plants without a nested RT-PCR detection.

Description

Technical field: [0001] The invention relates to a detection method of southern rice virus, in particular to a RT-PCR (reverse transcription polymerase chain reaction) detection method of southern rice black streaked dwarfvirus (SRBSDV). 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, Zhejiang Academy of Agricultural Sciences and South China Agricultural University reported that a new viral disease occurred in Guangdong and Hainan provinces respectively. South China Agricultural University tentatively named the virus as southern rice blackstreaked dwarf virus SRBSDV), and Zhejiang Academy of Agricultural Sciences called it rice black streaked dwarf virus 2 (rice black streaked dwarf virus 2, RBSDV2). In 2009, a dwarf disease similar to these two viral diseases occurred in a large area in Hunan Province. According to preliminary statistics, the affe...

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Problems solved by technology

[0003] Regarding the detection technology of SRBSDV, Zhou Guohui from South China Agricultural University and others designed two pairs of primers based on the nucleotide sequence of the tenth RNA segment of the virus for nested RT-PCR detection. This method requires 2 PCRs, which is time-consuming and consumable.

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