Department of Laboratory Methods and Medical Technology, Faculty of Health and Social Studies, University of South Bohemia, J Boreckeho 27, 370 11 Ceske Budejovice, Czech Republic.
J Virol Methods. 2010 Apr;165(1):64-70. doi: 10.1016/j.jviromet.2010.01.004. Epub 2010 Jan 25.
The success of DNA expression microarrays has been followed by applications of this technology to molecular diagnosis, mainly in the fields of biology and medicine. The experiments described below apply microarray diagnosis to agriculture. This report presents results of field tests for a DNA microarray designed to diagnose major viral potato pathogens. The assays were performed on samples that had been tested previously for the presence of viral infection by ELISA. RNA isolation methods were optimised for high sensitivity, using only 3 microg of total RNA that were reverse transcribed using random hexamers, with the resulting cDNA hybridised after labelling to an oligonucleotide array. The results obtained confirm the presence of pathogens indicated by ELISA and simultaneously reveal other viruses in the same reaction, showing that this method is appropriate for rapid detection of mixed viral infections. This observation was verified by subsequent RT-PCR and sequencing.
DNA 表达微阵列的成功应用，随后也被应用于分子诊断，主要在生物学和医学领域。下面描述的实验将微阵列诊断应用于农业。本报告介绍了一种用于诊断主要病毒性马铃薯病原体的 DNA 微阵列的现场测试结果。这些检测是在通过 ELISA 检测先前存在病毒感染的样本上进行的。使用仅 3 微克总 RNA 进行了 RNA 分离方法的优化，以提高灵敏度，使用随机六聚体进行逆转录，然后将所得 cDNA 进行标记并杂交到寡核苷酸阵列上。获得的结果证实了 ELISA 所示病原体的存在，并同时在同一反应中显示出其他病毒，表明该方法适用于快速检测混合病毒感染。这一观察结果随后通过 RT-PCR 和测序得到了验证。
