不同探针结构对应用微阵列技术检测兰花病毒结果的影响

Effect of different probe structures on the detection of orchid virus by microarray technology

根据黄瓜花叶病毒(CMV)、齿舌兰环斑病毒(ORSV)、建兰花叶病毒(Cym MV)、辣椒褪绿病毒(Ca CV)和落葵皱纹花叶病毒(Ba RMV)等5种危害较大的兰花病毒多重RT-PCR扩增产物序列,分别设计了5′端不同poly(d T)长短的探针,分别是:带poly(d T)15 spacer结构、带poly(d T)5 spacer结构、不带poly(d T)spacer结构,比较了3种探针紫外交联后的杂交效果。结果表明:5′端带poly(d T)15结构的探针杂交效率最高,而5′端不带poly(d T)结构的探针杂交效果极差,提示在探针设计时应选择在5′端带poly(d T)15结构以便获得更好的杂交效果。

According to the multiple RT-PCR amplification product sequences of the five more dangerous virus：Cucumber mosaic virus（CMV）,Odontoglossum ringspot virus（ORSV）,Cymbidium mosaic virus（CymMV）,Capsicum chlorosis virus（Ca CV）,Basella rugose mosaic virus（Ba RMV）,we designed probes with different length of 5′ end poly（d T）,respectively.The probes with the structure of poly（d T） 15 spacers,poly（d T） 5 spacers,and no poly（d T） spacer were used to compare the efficiency of hybridization after ultraviolet cross-linking. The results showed that the hybridization efficiency of probes with the structure of poly（d T） 15 spacers is the highest,while 5′ end without poly（d T） spacer is poor. Thus, probes with the structure of poly（d T） 15 spacers should be choose for better hybridization efficiency in the probe design.