Bystrická D, Lenz O, Mráz I, Dĕdic P, Síp M
Department of Plant Virology, Institute of Plant Molecular Biology, Academy of Sciences of the Czech Republic, Branisovská 31, 370 05 Ceské Budĕjovice, Czech Republic.
Acta Virol. 2003;47(1):41-4.
DNA microarray assay has become a useful tool for gene expression studies. Less frequent is its application to detection of viruses or diagnostics of virus diseases. Here we show design of a microscope slide-based microarray assay for simultaneous identification of several potato viruses. Different primer pairs were designed or adopted to obtain specific amplicons from six potato viruses: Potato virus A (PVA), Potato virus S (PVS), Potato virus X (PVX), Potato virus Y (PVY), Potato mop-top virus (PMTV) and Potato leaf-roll virus (PLRV). Purified viral DNA probes were spotted on a microscope slide coated with poly-L-lysine. The same primers were used for preparation of fluorochrome-labeled targets. The latter were denatured and hybridized on the microarray slide (chip). An example of simultaneous assay of two pathogens is given and possibilities of practical application of this type of assay are discussed.
DNA微阵列分析已成为基因表达研究的一种有用工具。其在病毒检测或病毒疾病诊断中的应用则较少见。在此,我们展示了一种基于显微镜载玻片的微阵列分析方法的设计,用于同时鉴定多种马铃薯病毒。设计或采用了不同的引物对,以从六种马铃薯病毒中获得特异性扩增子,这六种病毒分别是:马铃薯A病毒(PVA)、马铃薯S病毒(PVS)、马铃薯X病毒(PVX)、马铃薯Y病毒(PVY)、马铃薯帚顶病毒(PMTV)和马铃薯卷叶病毒(PLRV)。将纯化的病毒DNA探针点样在涂有聚-L-赖氨酸的显微镜载玻片上。使用相同的引物制备荧光染料标记的靶标。将后者变性并在微阵列载玻片(芯片)上进行杂交。给出了同时检测两种病原体的示例,并讨论了这种检测方法实际应用的可能性。