Species-independent detection of RNA virus by representational difference analysis using non-ribosomal hexanucleotides for reverse transcription.
Identifieur interne : 004620 ( Main/Exploration ); précédent : 004619; suivant : 004621Species-independent detection of RNA virus by representational difference analysis using non-ribosomal hexanucleotides for reverse transcription.
Auteurs : Daiji Endoh [Japon] ; Tetsuya Mizutani ; Rikio Kirisawa ; Yoshiyuki Maki ; Hidetoshi Saito ; Yasuhiro Kon ; Shigeru Morikawa ; Masanobu HayashiSource :
- Nucleic acids research [ 1362-4962 ] ; 2005.
Descripteurs français
- KwdFr :
- ADN complémentaire (), ADN complémentaire (biosynthèse), ARN ribosomique (), ARN viral (), ARN viral (analyse), Analyse de séquence d'ARN, Animaux, Bovins, Génome viral, Lignée cellulaire, Nucléotides (analyse), Rats, Réaction de polymérisation en chaîne (), Séquence nucléotidique, Transcription inverse, Virus du SRAS (génétique), Virus du SRAS (isolement et purification), Virus parainfluenza bovin de type 3 (génétique), Virus parainfluenza bovin de type 3 (isolement et purification), Virus à ARN (génétique), Virus à ARN (isolement et purification).
- MESH :
- analyse : ARN viral, Nucléotides.
- biosynthèse : ADN complémentaire.
- génétique : Virus du SRAS, Virus parainfluenza bovin de type 3, Virus à ARN.
- isolement et purification : Virus du SRAS, Virus parainfluenza bovin de type 3, Virus à ARN.
- ADN complémentaire, ARN ribosomique, ARN viral, Analyse de séquence d'ARN, Animaux, Bovins, Génome viral, Lignée cellulaire, Rats, Réaction de polymérisation en chaîne, Séquence nucléotidique, Transcription inverse.
English descriptors
- KwdEn :
- Animals, Base Sequence, Cattle, Cell Line, DNA, Complementary (biosynthesis), DNA, Complementary (chemistry), Genome, Viral, Nucleotides (analysis), Parainfluenza Virus 3, Bovine (genetics), Parainfluenza Virus 3, Bovine (isolation & purification), Polymerase Chain Reaction (methods), RNA Viruses (genetics), RNA Viruses (isolation & purification), RNA, Ribosomal (chemistry), RNA, Viral (analysis), RNA, Viral (chemistry), Rats, Reverse Transcription, SARS Virus (genetics), SARS Virus (isolation & purification), Sequence Analysis, RNA.
- MESH :
- chemical , analysis : Nucleotides, RNA, Viral.
- chemical , biosynthesis : DNA, Complementary.
- chemical , chemistry : DNA, Complementary, RNA, Ribosomal, RNA, Viral.
- genetics : Parainfluenza Virus 3, Bovine, RNA Viruses, SARS Virus.
- isolation & purification : Parainfluenza Virus 3, Bovine, RNA Viruses, SARS Virus.
- methods : Polymerase Chain Reaction.
- Animals, Base Sequence, Cattle, Cell Line, Genome, Viral, Rats, Reverse Transcription, Sequence Analysis, RNA.
Abstract
A method for the isolation of genomic fragments of RNA virus based on cDNA representational difference analysis (cDNA RDA) was developed. cDNA RDA has been applied for the subtraction of poly(A)(+) RNAs but not for poly(A)(-) RNAs, such as RNA virus genomes, owing to the vast quantity of ribosomal RNAs. We constructed primers for inefficient reverse transcription of ribosomal sequences based on the distribution analysis of hexanucleotide patterns in ribosomal RNA. The analysis revealed that distributions of hexanucleotide patterns in ribosomal RNA and virus genome were different. We constructed 96 hexanucleotides (non-ribosomal hexanucleotides) and used them as mixed primers for reverse transcription of cDNA RDA. A synchronous analysis of hexanucleotide patterns in known viral sequences showed that all the known genomic-size viral sequences include non-ribosomal hexanucleotides. In a model experiment, when non-ribosomal hexanucleotides were used as primers, in vitro transcribed plasmid RNA was efficiently reverse transcribed when compared with ribosomal RNA of rat cells. Using non-ribosomal primers, the cDNA fragments of severe acute respiratory syndrome coronavirus and bovine parainfluenza virus 3 were efficiently amplified by subtracting the cDNA amplicons derived from uninfected cells from those that were derived from virus-infected cells. The results suggest that cDNA RDA with non-ribosomal primers can be used for species-independent detection of viruses, including new viruses.
DOI: 10.1093/nar/gni064
PubMed: 15817564
Affiliations:
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Le document en format XML
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Rikio" sort="Kirisawa, Rikio" uniqKey="Kirisawa R" first="Rikio" last="Kirisawa">Rikio Kirisawa</name></author><author><name sortKey="Maki, Yoshiyuki" sort="Maki, Yoshiyuki" uniqKey="Maki Y" first="Yoshiyuki" last="Maki">Yoshiyuki Maki</name></author><author><name sortKey="Saito, Hidetoshi" sort="Saito, Hidetoshi" uniqKey="Saito H" first="Hidetoshi" last="Saito">Hidetoshi Saito</name></author><author><name sortKey="Kon, Yasuhiro" sort="Kon, Yasuhiro" uniqKey="Kon Y" first="Yasuhiro" last="Kon">Yasuhiro Kon</name></author><author><name sortKey="Morikawa, Shigeru" sort="Morikawa, Shigeru" uniqKey="Morikawa S" first="Shigeru" last="Morikawa">Shigeru Morikawa</name></author><author><name sortKey="Hayashi, Masanobu" sort="Hayashi, Masanobu" uniqKey="Hayashi M" first="Masanobu" last="Hayashi">Masanobu Hayashi</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2005">2005</date><idno type="RBID">pubmed:15817564</idno><idno 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hexanucleotides for reverse transcription.</title><author><name sortKey="Endoh, Daiji" sort="Endoh, Daiji" uniqKey="Endoh D" first="Daiji" last="Endoh">Daiji Endoh</name><affiliation wicri:level="1"><nlm:affiliation>Laboratory of Veterinary Radiology, School of Veterinary Medicine, Rakuno Gakuen University Ebetsu 069-8501, Japan. dendoh@rakuno.ac.jp</nlm:affiliation><country xml:lang="fr">Japon</country><wicri:regionArea>Laboratory of Veterinary Radiology, School of Veterinary Medicine, Rakuno Gakuen University Ebetsu 069-8501</wicri:regionArea><wicri:noRegion>Rakuno Gakuen University Ebetsu 069-8501</wicri:noRegion></affiliation></author><author><name sortKey="Mizutani, Tetsuya" sort="Mizutani, Tetsuya" uniqKey="Mizutani T" first="Tetsuya" last="Mizutani">Tetsuya Mizutani</name></author><author><name sortKey="Kirisawa, Rikio" sort="Kirisawa, Rikio" uniqKey="Kirisawa R" first="Rikio" last="Kirisawa">Rikio Kirisawa</name></author><author><name sortKey="Maki, Yoshiyuki" sort="Maki, 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Sequence</term><term>Cattle</term><term>Cell Line</term><term>DNA, Complementary (biosynthesis)</term><term>DNA, Complementary (chemistry)</term><term>Genome, Viral</term><term>Nucleotides (analysis)</term><term>Parainfluenza Virus 3, Bovine (genetics)</term><term>Parainfluenza Virus 3, Bovine (isolation & purification)</term><term>Polymerase Chain Reaction (methods)</term><term>RNA Viruses (genetics)</term><term>RNA Viruses (isolation & purification)</term><term>RNA, Ribosomal (chemistry)</term><term>RNA, Viral (analysis)</term><term>RNA, Viral (chemistry)</term><term>Rats</term><term>Reverse Transcription</term><term>SARS Virus (genetics)</term><term>SARS Virus (isolation & purification)</term><term>Sequence Analysis, RNA</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ADN complémentaire ()</term><term>ADN complémentaire (biosynthèse)</term><term>ARN ribosomique ()</term><term>ARN viral ()</term><term>ARN viral (analyse)</term><term>Analyse de séquence d'ARN</term><term>Animaux</term><term>Bovins</term><term>Génome viral</term><term>Lignée cellulaire</term><term>Nucléotides (analyse)</term><term>Rats</term><term>Réaction de polymérisation en chaîne ()</term><term>Séquence nucléotidique</term><term>Transcription inverse</term><term>Virus du SRAS (génétique)</term><term>Virus du SRAS (isolement et purification)</term><term>Virus parainfluenza bovin de type 3 (génétique)</term><term>Virus parainfluenza bovin de type 3 (isolement et purification)</term><term>Virus à ARN (génétique)</term><term>Virus à ARN (isolement et purification)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Nucleotides</term><term>RNA, Viral</term></keywords><keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>DNA, Complementary</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>DNA, Complementary</term><term>RNA, Ribosomal</term><term>RNA, Viral</term></keywords><keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>ARN viral</term><term>Nucléotides</term></keywords><keywords scheme="MESH" qualifier="biosynthèse" xml:lang="fr"><term>ADN complémentaire</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Parainfluenza Virus 3, Bovine</term><term>RNA Viruses</term><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Virus du SRAS</term><term>Virus parainfluenza bovin de type 3</term><term>Virus à ARN</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>Parainfluenza Virus 3, Bovine</term><term>RNA Viruses</term><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Virus du SRAS</term><term>Virus parainfluenza bovin de type 3</term><term>Virus à ARN</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Polymerase Chain Reaction</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Base Sequence</term><term>Cattle</term><term>Cell Line</term><term>Genome, Viral</term><term>Rats</term><term>Reverse Transcription</term><term>Sequence Analysis, RNA</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>ADN complémentaire</term><term>ARN ribosomique</term><term>ARN viral</term><term>Analyse de séquence d'ARN</term><term>Animaux</term><term>Bovins</term><term>Génome viral</term><term>Lignée cellulaire</term><term>Rats</term><term>Réaction de polymérisation en chaîne</term><term>Séquence nucléotidique</term><term>Transcription inverse</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A method for the isolation of genomic fragments of RNA virus based on cDNA representational difference analysis (cDNA RDA) was developed. cDNA RDA has been applied for the subtraction of poly(A)(+) RNAs but not for poly(A)(-) RNAs, such as RNA virus genomes, owing to the vast quantity of ribosomal RNAs. We constructed primers for inefficient reverse transcription of ribosomal sequences based on the distribution analysis of hexanucleotide patterns in ribosomal RNA. The analysis revealed that distributions of hexanucleotide patterns in ribosomal RNA and virus genome were different. We constructed 96 hexanucleotides (non-ribosomal hexanucleotides) and used them as mixed primers for reverse transcription of cDNA RDA. A synchronous analysis of hexanucleotide patterns in known viral sequences showed that all the known genomic-size viral sequences include non-ribosomal hexanucleotides. In a model experiment, when non-ribosomal hexanucleotides were used as primers, in vitro transcribed plasmid RNA was efficiently reverse transcribed when compared with ribosomal RNA of rat cells. Using non-ribosomal primers, the cDNA fragments of severe acute respiratory syndrome coronavirus and bovine parainfluenza virus 3 were efficiently amplified by subtracting the cDNA amplicons derived from uninfected cells from those that were derived from virus-infected cells. The results suggest that cDNA RDA with non-ribosomal primers can be used for species-independent detection of viruses, including new viruses.</div></front></TEI><affiliations><list><country><li>Japon</li></country></list><tree><noCountry><name sortKey="Hayashi, Masanobu" sort="Hayashi, Masanobu" uniqKey="Hayashi M" first="Masanobu" last="Hayashi">Masanobu Hayashi</name><name sortKey="Kirisawa, Rikio" sort="Kirisawa, Rikio" uniqKey="Kirisawa R" first="Rikio" last="Kirisawa">Rikio Kirisawa</name><name sortKey="Kon, Yasuhiro" sort="Kon, Yasuhiro" uniqKey="Kon Y" first="Yasuhiro" last="Kon">Yasuhiro Kon</name><name sortKey="Maki, Yoshiyuki" sort="Maki, Yoshiyuki" uniqKey="Maki Y" first="Yoshiyuki" last="Maki">Yoshiyuki Maki</name><name sortKey="Mizutani, Tetsuya" sort="Mizutani, Tetsuya" uniqKey="Mizutani T" first="Tetsuya" last="Mizutani">Tetsuya Mizutani</name><name sortKey="Morikawa, Shigeru" sort="Morikawa, Shigeru" uniqKey="Morikawa S" first="Shigeru" last="Morikawa">Shigeru Morikawa</name><name sortKey="Saito, Hidetoshi" sort="Saito, Hidetoshi" uniqKey="Saito H" first="Hidetoshi" last="Saito">Hidetoshi Saito</name></noCountry><country name="Japon"><noRegion><name sortKey="Endoh, Daiji" sort="Endoh, Daiji" uniqKey="Endoh D" first="Daiji" last="Endoh">Daiji Endoh</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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