Identification of dengue sequences by genomic amplification: rapid diagnosis of dengue virus serotypes in peripheral blood
Polymerase chain reaction (PCR) was developed for the in vitro amplification of dengue virus RNA via cDNA. A fraction of the N-terminus gene of the envelope protein in the four dengue serotypes was amplified using synthetic oligonucleotide primer pairs. Amplified products were cloned and used as den...
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| Veröffentlicht in: | Journal of virological methods 1990-10, Vol.30 (1), p.41-54 |
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| container_title | Journal of virological methods |
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| creator | Deubel, Vincent Laille, Manola Hugnot, Jean-Philippe Chungue, Eliane Guesdon, Jean-Luc Drouet, Marie Thérèse Bassot, Sylviane Chevrier, Daniele |
| description | Polymerase chain reaction (PCR) was developed for the in vitro amplification of dengue virus RNA via cDNA. A fraction of the N-terminus gene of the envelope protein in the four dengue serotypes was amplified using synthetic oligonucleotide primer pairs. Amplified products were cloned and used as dengue type-specific probes in gel electrophoresis and dot-blot hybridization. We detected and characterized dengue virus serotypes in blood samples by the three-step procedure DNA-PAH consisting in cDNA priming (P), DNA amplification (A) and hybridization (H) using specific non-radiolabelled probes. Our findings showed that DNA-PAH was more rapid and sensitive in the identification of the infecting serotype than the mosquito cell cultures. Moreover, the failure of cultures to detect virus particles in sera containing few copies of viral genome or anti-dengue antibodies justified the approach of DNA-PAH to the dengue identification in clinical specimens. |
| doi_str_mv | 10.1016/0166-0934(90)90042-E |
| format | Article |
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A fraction of the N-terminus gene of the envelope protein in the four dengue serotypes was amplified using synthetic oligonucleotide primer pairs. Amplified products were cloned and used as dengue type-specific probes in gel electrophoresis and dot-blot hybridization. We detected and characterized dengue virus serotypes in blood samples by the three-step procedure DNA-PAH consisting in cDNA priming (P), DNA amplification (A) and hybridization (H) using specific non-radiolabelled probes. Our findings showed that DNA-PAH was more rapid and sensitive in the identification of the infecting serotype than the mosquito cell cultures. Moreover, the failure of cultures to detect virus particles in sera containing few copies of viral genome or anti-dengue antibodies justified the approach of DNA-PAH to the dengue identification in clinical specimens.</description><identifier>ISSN: 0166-0934</identifier><identifier>EISSN: 1879-0984</identifier><identifier>DOI: 10.1016/0166-0934(90)90042-E</identifier><identifier>PMID: 2086596</identifier><identifier>CODEN: JVMEDH</identifier><language>eng</language><publisher>London: Elsevier B.V</publisher><subject>Base Sequence ; Biological and medical sciences ; Blotting, Northern - methods ; Blotting, Southern - methods ; Dengue - blood ; Dengue virus ; Dengue Virus - classification ; Dengue Virus - genetics ; Dengue Virus - isolation & purification ; Fundamental and applied biological sciences. Psychology ; Genes, Viral ; Humans ; Microbiology ; Molecular Sequence Data ; Nucleic Acid Hybridization ; Oligonucleotide Probes ; Polymerase Chain Reaction - methods ; Polynucleotide chain reaction ; RNA, Viral - genetics ; RNA, Viral - isolation & purification ; Serotyping ; Techniques used in virology ; Virology ; Virus diagnosis</subject><ispartof>Journal of virological methods, 1990-10, Vol.30 (1), p.41-54</ispartof><rights>1990</rights><rights>1991 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c418t-51ead1af22d77bd3a22e10fb6c4a5a383319935483dbf3415571aba565aa18b83</citedby><cites>FETCH-LOGICAL-c418t-51ead1af22d77bd3a22e10fb6c4a5a383319935483dbf3415571aba565aa18b83</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/0166-0934(90)90042-E$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,778,782,3539,27911,27912,45982</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=19392862$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/2086596$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Deubel, Vincent</creatorcontrib><creatorcontrib>Laille, Manola</creatorcontrib><creatorcontrib>Hugnot, Jean-Philippe</creatorcontrib><creatorcontrib>Chungue, Eliane</creatorcontrib><creatorcontrib>Guesdon, Jean-Luc</creatorcontrib><creatorcontrib>Drouet, Marie Thérèse</creatorcontrib><creatorcontrib>Bassot, Sylviane</creatorcontrib><creatorcontrib>Chevrier, Daniele</creatorcontrib><title>Identification of dengue sequences by genomic amplification: rapid diagnosis of dengue virus serotypes in peripheral blood</title><title>Journal of virological methods</title><addtitle>J Virol Methods</addtitle><description>Polymerase chain reaction (PCR) was developed for the in vitro amplification of dengue virus RNA via cDNA. A fraction of the N-terminus gene of the envelope protein in the four dengue serotypes was amplified using synthetic oligonucleotide primer pairs. Amplified products were cloned and used as dengue type-specific probes in gel electrophoresis and dot-blot hybridization. We detected and characterized dengue virus serotypes in blood samples by the three-step procedure DNA-PAH consisting in cDNA priming (P), DNA amplification (A) and hybridization (H) using specific non-radiolabelled probes. Our findings showed that DNA-PAH was more rapid and sensitive in the identification of the infecting serotype than the mosquito cell cultures. Moreover, the failure of cultures to detect virus particles in sera containing few copies of viral genome or anti-dengue antibodies justified the approach of DNA-PAH to the dengue identification in clinical specimens.</description><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Blotting, Northern - methods</subject><subject>Blotting, Southern - methods</subject><subject>Dengue - blood</subject><subject>Dengue virus</subject><subject>Dengue Virus - classification</subject><subject>Dengue Virus - genetics</subject><subject>Dengue Virus - isolation & purification</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genes, Viral</subject><subject>Humans</subject><subject>Microbiology</subject><subject>Molecular Sequence Data</subject><subject>Nucleic Acid Hybridization</subject><subject>Oligonucleotide Probes</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Polynucleotide chain reaction</subject><subject>RNA, Viral - genetics</subject><subject>RNA, Viral - isolation & purification</subject><subject>Serotyping</subject><subject>Techniques used in virology</subject><subject>Virology</subject><subject>Virus diagnosis</subject><issn>0166-0934</issn><issn>1879-0984</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1990</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkV-L1DAUxYO4rOPoN1DIi6IP1fxp0sQHQZbRXVjwRZ_DbZKOkbapSbswfvpNnWH0yX0IgXt_55Ccg9ALSt5RQuX7cmRFNK_faPJWE1KzavcIbahqdBmr-jHanJEn6GnOPwkhouH8El0yoqTQcoN-3zg_zqELFuYQRxw7XAb7xePsfy1-tD7j9oD3foxDsBiGqT_DH3CCKTjsAuzHmEP-R30X0pKLR4rzYSoeYcSTT2H64RP0uO1jdM_QRQd99s9P9xZ9_7z7dnVd3X79cnP16bayNVVzJagHR6FjzDVN6zgw5inpWmlrEMAV51RrLmrFXdvxmgrRUGhBSAFAVav4Fr0--k4pli_l2QwhW9_3MPq4ZKMIp1Ix_iBIJW30GuAW1UfQpphz8p2ZUhggHQwlZu3GrMGbNXijifnTjdkV2cuT_9IO3p1FpzLK_tVpD9lC3yUYbch_vTXXTElWuI9HzpfU7oJPJtuwVuVC8nY2Lob_P-QejdSsuQ</recordid><startdate>19901001</startdate><enddate>19901001</enddate><creator>Deubel, Vincent</creator><creator>Laille, Manola</creator><creator>Hugnot, Jean-Philippe</creator><creator>Chungue, Eliane</creator><creator>Guesdon, Jean-Luc</creator><creator>Drouet, Marie Thérèse</creator><creator>Bassot, Sylviane</creator><creator>Chevrier, Daniele</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7U9</scope><scope>H94</scope><scope>7X8</scope></search><sort><creationdate>19901001</creationdate><title>Identification of dengue sequences by genomic amplification: rapid diagnosis of dengue virus serotypes in peripheral blood</title><author>Deubel, Vincent ; Laille, Manola ; Hugnot, Jean-Philippe ; Chungue, Eliane ; Guesdon, Jean-Luc ; Drouet, Marie Thérèse ; Bassot, Sylviane ; Chevrier, Daniele</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c418t-51ead1af22d77bd3a22e10fb6c4a5a383319935483dbf3415571aba565aa18b83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1990</creationdate><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Blotting, Northern - methods</topic><topic>Blotting, Southern - methods</topic><topic>Dengue - blood</topic><topic>Dengue virus</topic><topic>Dengue Virus - classification</topic><topic>Dengue Virus - genetics</topic><topic>Dengue Virus - isolation & purification</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genes, Viral</topic><topic>Humans</topic><topic>Microbiology</topic><topic>Molecular Sequence Data</topic><topic>Nucleic Acid Hybridization</topic><topic>Oligonucleotide Probes</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Polynucleotide chain reaction</topic><topic>RNA, Viral - genetics</topic><topic>RNA, Viral - isolation & purification</topic><topic>Serotyping</topic><topic>Techniques used in virology</topic><topic>Virology</topic><topic>Virus diagnosis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Deubel, Vincent</creatorcontrib><creatorcontrib>Laille, Manola</creatorcontrib><creatorcontrib>Hugnot, Jean-Philippe</creatorcontrib><creatorcontrib>Chungue, Eliane</creatorcontrib><creatorcontrib>Guesdon, Jean-Luc</creatorcontrib><creatorcontrib>Drouet, Marie Thérèse</creatorcontrib><creatorcontrib>Bassot, Sylviane</creatorcontrib><creatorcontrib>Chevrier, Daniele</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of virological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Deubel, Vincent</au><au>Laille, Manola</au><au>Hugnot, Jean-Philippe</au><au>Chungue, Eliane</au><au>Guesdon, Jean-Luc</au><au>Drouet, Marie Thérèse</au><au>Bassot, Sylviane</au><au>Chevrier, Daniele</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Identification of dengue sequences by genomic amplification: rapid diagnosis of dengue virus serotypes in peripheral blood</atitle><jtitle>Journal of virological methods</jtitle><addtitle>J Virol Methods</addtitle><date>1990-10-01</date><risdate>1990</risdate><volume>30</volume><issue>1</issue><spage>41</spage><epage>54</epage><pages>41-54</pages><issn>0166-0934</issn><eissn>1879-0984</eissn><coden>JVMEDH</coden><abstract>Polymerase chain reaction (PCR) was developed for the in vitro amplification of dengue virus RNA via cDNA. A fraction of the N-terminus gene of the envelope protein in the four dengue serotypes was amplified using synthetic oligonucleotide primer pairs. Amplified products were cloned and used as dengue type-specific probes in gel electrophoresis and dot-blot hybridization. We detected and characterized dengue virus serotypes in blood samples by the three-step procedure DNA-PAH consisting in cDNA priming (P), DNA amplification (A) and hybridization (H) using specific non-radiolabelled probes. Our findings showed that DNA-PAH was more rapid and sensitive in the identification of the infecting serotype than the mosquito cell cultures. 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| ispartof | Journal of virological methods, 1990-10, Vol.30 (1), p.41-54 |
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| subjects | Base Sequence Biological and medical sciences Blotting, Northern - methods Blotting, Southern - methods Dengue - blood Dengue virus Dengue Virus - classification Dengue Virus - genetics Dengue Virus - isolation & purification Fundamental and applied biological sciences. Psychology Genes, Viral Humans Microbiology Molecular Sequence Data Nucleic Acid Hybridization Oligonucleotide Probes Polymerase Chain Reaction - methods Polynucleotide chain reaction RNA, Viral - genetics RNA, Viral - isolation & purification Serotyping Techniques used in virology Virology Virus diagnosis |
| title | Identification of dengue sequences by genomic amplification: rapid diagnosis of dengue virus serotypes in peripheral blood |
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