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Rotavirus gene detection with biotinylated single-stranded RNA probes.

Abstract：

:Biotinylated single-stranded RNA probes from two of the eleven genome segments of the simian rotavirus SA11 were synthesized from cloned DNA and used in dot-blot and Northern-blot hybridization assays. Different types of membranes and conditions to prepare and use synthetic non-radioactive transcript probes were evaluated to obtain optimal test results. Nytran membranes showed the highest sensitivity and lowest backgrounds for hybridization with biotinylated RNA probes. When a gene 6 single-stranded biotinylated probe was used in a dot-blot format, test sensitivity was 0.1 ng for detection of homologous RNA and 0.4-1.5 micrograms for detection of RNA from heterologous rotavirus strains. When used in Northern blots, detection with this gene 6 probe required 1 ng of total SA11RNA or 50 ng of heterologous RNA to be applied to the gels for transfer. Simultaneous hybridization with probes from two different genes on one membrane showed a detection level similar to that seen with single probes alone. The advantages of using biotinylated single-stranded RNA probes to detect or characterize the genes of viruses with double-stranded RNA genomes are shown.

journal_name

Mol Cell Probes

journal_title

Molecular and cellular probes

authors

Bellinzoni R,Xi JA,Tanaka TN,Scodeller E,Estes MK

doi

10.1016/0890-8508(89)90004-2

subject

Has Abstract

pub_date

1989-09-01 00:00:00

pages

233-44

issue

3

eissn

0890-8508

issn

1096-1194

journal_volume

3

pub_type

杂志文章

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