Enzymatic amplification of latent pseudorabies virus nucleic acid sequences
Identifieur interne : 001E81 ( Istex/Checkpoint ); précédent : 001E80; suivant : 001E82Enzymatic amplification of latent pseudorabies virus nucleic acid sequences
Auteurs : J. R Lokensgard [États-Unis] ; D. G. Thawley [États-Unis] ; T. W. Molitor [États-Unis]Source :
- Journal of Virological Methods [ 0166-0934 ] ; 1991.
English descriptors
- Teeft :
- Acute phase, Amplification, Amplification techniques, Antisense orientation, Assay, Bamhl fragment, Buffer control, Cells harbor, Cheung, Coding strand, Denatured ssdna, Denhardt solution, Enzymatic amplification, Ethidium bromide, Ganglion, Gene expression, Genome, Hybridization, Internal oligonucleotide, Internal probe, Latency, Latent pseudorabies virus, Latent state, Latently, Nucleic, Nucleic acid amplification methods, Nucleic acids, Oligonucleotide, Polymerase, Polymerase chain reaction, Positive control, Primer, Pseudorabies, Pseudorabies virus, Pseudorabies virus genome, Room temperature, Saint paul, Same polarity, Sequence data, Sodium phosphate, Standard dntps, Swine, Template strand, Trigeminal ganglia, Uninfected swine, Vero cells, Viral, Viral gene expression, Virol.
Abstract
Abstract: To investigate various aspects of the latency of pseudorabies virus in swine (PRV, suid herpesvirus 1) we developed in vitro nucleic acid amplification methods based upon the polymerase chain reaction. Primers flanking a 156-bp region of the pseudorabies virus gp II gene were annealed to purified PRV DNA as well as DNA isolated from the trigeminal ganglia of swine latently infected with PRV and subjected to PCR amplification. Following amplification, 100 fg of PRV DNA was visualizable on stained gels and 1 fg (equivalent to 6 viral genome copies) was detectable when amplification was combined with blot hybridization. PRV-specific DNA sequences which remained undetectable by direct blot hybridization assays were amplified to levels visualizable on ethidium-bromide-stained gels in 5 of 5 experimental latently infected animals. In addition, oligonucleotide primers specific for a 223-bp region of the PRV immediate-early gene (IE 180) were capable of amplifying overlapping latency associated transcripts (LATs), via a cDNA intermediate, in 6 of 6 latently infected swine. These nucleic acid amplification methods should be applicable to the investigation of PRV latency, and gene expression during latency and reactivation, in which few cells harbor latent virus.
Url:
DOI: 10.1016/0166-0934(91)90120-O
Affiliations:
Links toward previous steps (curation, corpus...)
Links to Exploration step
ISTEX:1D6488862892600F56DB36C7AEB7DC76FD838D0ALe document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title>Enzymatic amplification of latent pseudorabies virus nucleic acid sequences</title><author><name sortKey="Lokensgard, J R" sort="Lokensgard, J R" uniqKey="Lokensgard J" first="J. R" last="Lokensgard">J. R Lokensgard</name></author><author><name sortKey="Thawley, D G" sort="Thawley, D G" uniqKey="Thawley D" first="D. G." last="Thawley">D. G. Thawley</name></author><author><name sortKey="Molitor, T W" sort="Molitor, T W" uniqKey="Molitor T" first="T. W." last="Molitor">T. W. Molitor</name></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:1D6488862892600F56DB36C7AEB7DC76FD838D0A</idno><date when="1991" year="1991">1991</date><idno type="doi">10.1016/0166-0934(91)90120-O</idno><idno type="url">https://api.istex.fr/ark:/67375/6H6-WPWQB8GC-R/fulltext.pdf</idno><idno type="wicri:Area/Istex/Corpus">001033</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">001033</idno><idno type="wicri:Area/Istex/Curation">001033</idno><idno type="wicri:Area/Istex/Checkpoint">001E81</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Checkpoint">001E81</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a">Enzymatic amplification of latent pseudorabies virus nucleic acid sequences</title><author><name sortKey="Lokensgard, J R" sort="Lokensgard, J R" uniqKey="Lokensgard J" first="J. R" last="Lokensgard">J. R Lokensgard</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Clinical and Population Sciences, College of Veterinary Medicine, University of Minnesota, Saint Paul, Minnesota</wicri:regionArea><placeName><region type="state">Minnesota</region></placeName></affiliation></author><author><name sortKey="Thawley, D G" sort="Thawley, D G" uniqKey="Thawley D" first="D. G." last="Thawley">D. G. Thawley</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Clinical and Population Sciences, College of Veterinary Medicine, University of Minnesota, Saint Paul, Minnesota</wicri:regionArea><placeName><region type="state">Minnesota</region></placeName></affiliation></author><author><name sortKey="Molitor, T W" sort="Molitor, T W" uniqKey="Molitor T" first="T. W." last="Molitor">T. W. Molitor</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Clinical and Population Sciences, College of Veterinary Medicine, University of Minnesota, Saint Paul, Minnesota</wicri:regionArea><placeName><region type="state">Minnesota</region></placeName></affiliation></author></analytic><monogr></monogr><series><title level="j">Journal of Virological Methods</title><title level="j" type="abbrev">VIRMET</title><idno type="ISSN">0166-0934</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1991">1991</date><biblScope unit="volume">34</biblScope><biblScope unit="issue">1</biblScope><biblScope unit="page" from="45">45</biblScope><biblScope unit="page" to="55">55</biblScope></imprint><idno type="ISSN">0166-0934</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0166-0934</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Acute phase</term><term>Amplification</term><term>Amplification techniques</term><term>Antisense orientation</term><term>Assay</term><term>Bamhl fragment</term><term>Buffer control</term><term>Cells harbor</term><term>Cheung</term><term>Coding strand</term><term>Denatured ssdna</term><term>Denhardt solution</term><term>Enzymatic amplification</term><term>Ethidium bromide</term><term>Ganglion</term><term>Gene expression</term><term>Genome</term><term>Hybridization</term><term>Internal oligonucleotide</term><term>Internal probe</term><term>Latency</term><term>Latent pseudorabies virus</term><term>Latent state</term><term>Latently</term><term>Nucleic</term><term>Nucleic acid amplification methods</term><term>Nucleic acids</term><term>Oligonucleotide</term><term>Polymerase</term><term>Polymerase chain reaction</term><term>Positive control</term><term>Primer</term><term>Pseudorabies</term><term>Pseudorabies virus</term><term>Pseudorabies virus genome</term><term>Room temperature</term><term>Saint paul</term><term>Same polarity</term><term>Sequence data</term><term>Sodium phosphate</term><term>Standard dntps</term><term>Swine</term><term>Template strand</term><term>Trigeminal ganglia</term><term>Uninfected swine</term><term>Vero cells</term><term>Viral</term><term>Viral gene expression</term><term>Virol</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: To investigate various aspects of the latency of pseudorabies virus in swine (PRV, suid herpesvirus 1) we developed in vitro nucleic acid amplification methods based upon the polymerase chain reaction. Primers flanking a 156-bp region of the pseudorabies virus gp II gene were annealed to purified PRV DNA as well as DNA isolated from the trigeminal ganglia of swine latently infected with PRV and subjected to PCR amplification. Following amplification, 100 fg of PRV DNA was visualizable on stained gels and 1 fg (equivalent to 6 viral genome copies) was detectable when amplification was combined with blot hybridization. PRV-specific DNA sequences which remained undetectable by direct blot hybridization assays were amplified to levels visualizable on ethidium-bromide-stained gels in 5 of 5 experimental latently infected animals. In addition, oligonucleotide primers specific for a 223-bp region of the PRV immediate-early gene (IE 180) were capable of amplifying overlapping latency associated transcripts (LATs), via a cDNA intermediate, in 6 of 6 latently infected swine. These nucleic acid amplification methods should be applicable to the investigation of PRV latency, and gene expression during latency and reactivation, in which few cells harbor latent virus.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Minnesota</li></region></list><tree><country name="États-Unis"><region name="Minnesota"><name sortKey="Lokensgard, J R" sort="Lokensgard, J R" uniqKey="Lokensgard J" first="J. R" last="Lokensgard">J. R Lokensgard</name></region><name sortKey="Molitor, T W" sort="Molitor, T W" uniqKey="Molitor T" first="T. W." last="Molitor">T. W. Molitor</name><name sortKey="Thawley, D G" sort="Thawley, D G" uniqKey="Thawley D" first="D. G." last="Thawley">D. G. Thawley</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/MersV1/Data/Istex/Checkpoint
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 001E81 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Istex/Checkpoint/biblio.hfd -nk 001E81 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= MersV1
|flux= Istex
|étape= Checkpoint
|type= RBID
|clé= ISTEX:1D6488862892600F56DB36C7AEB7DC76FD838D0A
|texte= Enzymatic amplification of latent pseudorabies virus nucleic acid sequences
}}
| This area was generated with Dilib version V0.6.33. |  |