RT-PCR检测齿兰环斑病毒技术的建立

Development of RT-PCR assay for the detection of Odntoglossum rongspot virus

根据已报道的齿兰环斑病毒(ORSV)外壳蛋白(CP)基因序列,进行同源性比较,设计了一对各为20bp的引物.通过优化试验条件,建立了稳定的RT-PCR检测ORSV体系,其检测灵敏度可达0.01ng·mL-1.将此方法与ELISA进行比较,结果表明,RT-PCR检测灵敏度比ELISA高1000倍.应用这两种方法同时检测了138瓶兰花组培苗样品,其中RT-PCR检测出32瓶兰花组培苗感染病毒,而ELISA只能检测其中的18瓶.

Using a pair of 20 bp primers complementary to the conserved Odntoglossum ringspot virus (ORSV) coat protein (CP) gene sequences published previously, the sensitive RTPCR method was set up to test ORSV immediately by optimizing experiment conditions. The sensitivity of RTPCR method can come to 0.01 ng·mL-1 ORSV, which is 1000 times higher than ELISA. And 138 samples suspected to be infected with ORSV were tested by both of the two methods. The results showed that 32 of the samples were positive by RTPCR, but only 18 were positive by ELISA.