根据黄瓜绿斑驳花叶病毒不同分离物外壳蛋白基因(CP)的保守序列,设计合成引物和TaqMan荧光探针,建立了黄瓜绿斑驳花叶病毒的实时荧光RT-PCR(Real time RT-PCR)检测方法。该方法利用TaqMan探针水解产生的荧光信号实时监测目标基因的扩增,实现PCR扩增与检测同步进行。结果表明,实时荧光RT-PCR方法检测灵敏度比普通RT-PCR高约10倍,并且具有快速、简便、准确的优点,适合于CGMMV的快速、高效检测。
The nucleotide sequence of small coat protein(CPS) gene of Broad bean stain virus(BBSV) was determined and compared with other comoviruses.The CPS gene of BBSV consisted of 687 nucleotides and encodes a putative protein of 228 amino acid residues.The CPS sequence of BBSV and those of other comoviruses shared identities of 36.5%-58.9% and 35.2%-70.3% at the nucleotide and amino acid levels,respectively.The RT-PCR method specific for BBSV detection was developed based on the determined CPS sequence.The RT-PCR assay presented here allows,for the first time,rapid and specific detection of BBSV.
