Linked Life Data

14600342

Source:http://linkedlifedata.com/resource/pubmed/id/14600342

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
10
pubmed:dateCreated
2003-11-5
pubmed:abstractText
A sensitive and specific in situ amplification technique is needed to elucidate the dynamics of rabies virus in the body during the long incubation period after infection. To overcome the disadvantage of using the traditional reverse transcription (RT)-PCR in in situ studies, an isothermal nucleic acid sequence-based amplification (NASBA) technique was developed for detection of the rabies virus gene. The NASBA technique involves the use of 4 enzymatic activities to produce multiple RNA copies of the target sequence by means of double-strand cDNA intermediates under an isothermal condition without thermocycling. The amplified cDNA intermediates from the genomic RNA in the rabies virion and the total RNA in the infected cells in NASBA reaction were analyzed by Southern hybridization assays. The specific amplified products of the rabies viral gene with the expected length were detected after 8 hr of incubation in NASBA using both of the RNAs as templates. The NASBA system used in this study was less sensitive than the general RT-PCR technique. This may have been because we employed Southern hybridization for the amplified cDNA intermediates, not many RNA copies, to evaluate the NASBA results. In conclusion, we successfully amplified the rabies viral gene in the NASBA reaction under an isothermal condition. The unique character of this technique would make it particularly valuable for in situ studies not only on rabies virus but also on other RNA viruses.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
0916-7250
pubmed:author
pubmed:issnType
Print
pubmed:volume
65
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1063-8
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed:year
2003
pubmed:articleTitle
Isothermal amplification of rabies virus gene.
pubmed:affiliation
Laboratory of Zoonotic Diseases, Faculty of Agriculture, Gifu University, Yanagido, Gifu, Japan.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't