中国生物工程杂志

Based on the conserved region nucleotide sequences of four viruses (Alfalfa Mosaic Virus, ALMV; Cucumber mosaic virus, CMV; Cucumber mosaic virus-satellite, CMV-sat; Potato virus Y, PVY) and one viroid (Potato spindle tuber viroid, PSTVd) which could be pathogenic on potato world and one inner control(18S rRNA), specific oligonucleotide probes and primers were designed and spotting for hybridization detection.There is examine the influence of probe concentration、hybridization time、hybridization temperature and spotting solutions on microarray signal.Finally there is validate the specificity of optimized plant virus detection array.The experiment indication: There is not significant influence on hybridization signals with the concentration of the oligo probes between 5-20 µM,PCR probe show the linear relationship in influence of hybridization signals. Microarray hybridization at 45℃ for 4 h, the intensity of hybridization signals was the most significant, and under this condition the influence tendency on oligonucleotide probes and PCR probes was nearly same. With the comparison of different spotting solutions, DMSO had the best spotting reproducibility. Through the whole condition optimized, the oligonucleotide probe microarray and PCR probe microarray were provided with better specificity in the hybridized detection.