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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。

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Biotinylated nucleic acid hybridization probes for potato virus detection.用于马铃薯病毒检测的生物素化核酸杂交探针。
Arch Virol. 1988;103(3-4):231-41. doi: 10.1007/BF01311095.
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本文引用的文献

1
Sensitive and rapid diagnosis of potato spindle tuber viroid disease by nucleic Acid hybridization.利用核酸杂交技术对马铃薯纺锤块茎类病毒病进行灵敏快速诊断。
Science. 1981 Aug 7;213(4508):670-2. doi: 10.1126/science.213.4508.670.
2
The application of spot hybridization to the detection of DNA and RNA viruses in plant tissues.斑点杂交在植物组织中DNA和RNA病毒检测中的应用。
J Virol Methods. 1983 Apr;6(4):215-24. doi: 10.1016/0166-0934(83)90048-4.
3
Chemically modified nucleic acids as immunodetectable probes in hybridization experiments.
化学修饰的核酸作为杂交实验中可免疫检测的探针。
Proc Natl Acad Sci U S A. 1984 Jun;81(11):3466-70. doi: 10.1073/pnas.81.11.3466.
4
A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity.一种将DNA限制性内切酶片段放射性标记至高比活度的技术。
Anal Biochem. 1983 Jul 1;132(1):6-13. doi: 10.1016/0003-2697(83)90418-9.
5
A simple and very efficient method for generating cDNA libraries.一种简单且非常有效的生成cDNA文库的方法。
Gene. 1983 Nov;25(2-3):263-9. doi: 10.1016/0378-1119(83)90230-5.
6
Rapid and sensitive colorimetric method for visualizing biotin-labeled DNA probes hybridized to DNA or RNA immobilized on nitrocellulose: Bio-blots.用于可视化与固定在硝酸纤维素膜上的DNA或RNA杂交的生物素标记DNA探针的快速灵敏比色法:生物印迹法。
Proc Natl Acad Sci U S A. 1983 Jul;80(13):4045-9. doi: 10.1073/pnas.80.13.4045.
7
Sandwich nucleic acid hybridization: a method with a universally usable labeled probe for various specific tests.夹心核酸杂交:一种用于各种特异性检测的具有通用标记探针的方法。
J Virol Methods. 1986 Apr;13(1):1-8. doi: 10.1016/0166-0934(86)90066-2.
8
Nonradioactive, photobiotin-labelled DNA probes for the routine diagnosis of barley yellow dwarf virus.
J Virol Methods. 1987 Jun;16(3):225-37. doi: 10.1016/0166-0934(87)90007-3.
9
Detection of sub-picogram quantities of specific DNA sequences on blot hybridization with biotinylated probes.用生物素化探针进行印迹杂交时检测亚皮克级特定DNA序列量。
Nucleic Acids Res. 1985 Nov 25;13(22):8083-91. doi: 10.1093/nar/13.22.8083.
10
Non-radioactive hybridization probes prepared by the chemical labelling of DNA and RNA with a novel reagent, photobiotin.通过用新型试剂光生物素对DNA和RNA进行化学标记制备的非放射性杂交探针。
Nucleic Acids Res. 1985 Feb 11;13(3):745-61. doi: 10.1093/nar/13.3.745.