用于马铃薯病毒检测的生物素化核酸杂交探针。

Biotinylated nucleic acid hybridization probes for potato virus detection.

作者信息

Hopp H E, Giavedoni L, Mandel M A, Arese A, Orman B, Bravo Almonacid F, Torres H N, Mentaberry A N

机构信息

Instituto de Biologiá Molecular CICV-INTA Castelar, Buenos Aires, Argentina.

出版信息

Arch Virol. 1988;103(3-4):231-41. doi: 10.1007/BF01311095.

DOI:10.1007/BF01311095

PMID:3214274

Abstract

cDNA libraries, representative of potato viruses X (PVXc strain) and Y (PVY degrees strain) genomes were obtained. A PVX cDNA cloned fragment was sequenced and biotinylated to be used as hybridization probe for the detection of purified virus or nucleic acid extracts of infected plants. Dot hybridization assay was sensitive to detect 4 ng of viral particles, corresponding to about 200 pg of viral RNA. The level of detection in infected plant extracts was as effective as that obtained with the ELISA. The presence of biotinylated PVY cDNA in the hybridization mixture did not affect sensitivity of the PVX detection assay, suggesting that a single diagnostic assay for several potato viruses and virus-related pathogens could be developed.

摘要

获得了代表马铃薯X病毒（PVXc株系）和Y病毒（PVY°株系）基因组的cDNA文库。对一个PVX cDNA克隆片段进行了测序并进行生物素化处理，用作杂交探针，用于检测纯化病毒或受感染植物的核酸提取物。斑点杂交试验检测4 ng病毒颗粒很灵敏，这相当于约200 pg病毒RNA。在受感染植物提取物中的检测水平与酶联免疫吸附测定法的检测水平一样有效。杂交混合物中生物素化的PVY cDNA的存在并不影响PVX检测试验的灵敏度，这表明可以开发一种针对多种马铃薯病毒和病毒相关病原体的单一诊断试验。