Genosensor on gold films with enzymatic electrochemical detection of a SARS virus sequence☆
Identifieur interne : 000E58 ( Pmc/Curation ); précédent : 000E57; suivant : 000E59Genosensor on gold films with enzymatic electrochemical detection of a SARS virus sequence☆
Auteurs : Patricia Abad-Valle ; M. Teresa Fernández-Abedul ; Agustín Costa-GarcíaSource :
- Biosensors & Bioelectronics [ 0956-5663 ] ; 2004.
Abstract
A hybridisation-based genosensor was designed on a 100 nm sputtered gold film. This material worked as an immobilisation and transduction surface. A 30-mer sequence that encodes a short lysine-rich region, unique to SARS (severe acute respiratory syndrome) virus, was chosen as target. A complementary strand (probe), labelled with a thiol group at the 3′-end, was immobilised on the film. After blocking the surface, hybridisation with the biotin-conjugated SARS strand (at the 3′-end) took place. Interaction with alkaline phosphatase-labelled streptavidin permits amplified indirect electrochemical detection. The analytical signal is constituted by an electrochemical process of indigo carmine, the soluble product of the enzymatic hydrolysis of 3-indoxyl phosphate. The use of a sensitive electrochemical technique such as square wave voltammetry allowed a detection limit of 6 pM to be obtained for this DNA sequence, lower than any other found in the bibliography. The parameters affecting the methodology were studied, with special attention being placed on selectivity. Specificity was clearly enhanced when interaction time and stringency (in the form of formamide percentage) were increased. With 1 h of strand interaction and employing 50% of formamide in the hybridisation buffer, a 3-base mismatch strand was perfectly distinguished from the complementary.
Url:
DOI: 10.1016/j.bios.2004.10.019
PubMed: 15797323
PubMed Central: 7126974
Links toward previous steps (curation, corpus...)
- to stream Pmc, to step Corpus: Pour aller vers cette notice dans l'étape Curation :000E58
Links to Exploration step
PMC:7126974Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Genosensor on gold films with enzymatic electrochemical detection of a SARS virus sequence<sup><xref ref-type="fn" rid="d32e1184">☆</xref></sup></title><author><name sortKey="Abad Valle, Patricia" sort="Abad Valle, Patricia" uniqKey="Abad Valle P" first="Patricia" last="Abad-Valle">Patricia Abad-Valle</name></author><author><name sortKey="Fernandez Abedul, M Teresa" sort="Fernandez Abedul, M Teresa" uniqKey="Fernandez Abedul M" first="M. Teresa" last="Fernández-Abedul">M. Teresa Fernández-Abedul</name></author><author><name sortKey="Costa Garcia, Agustin" sort="Costa Garcia, Agustin" uniqKey="Costa Garcia A" first="Agustín" last="Costa-García">Agustín Costa-García</name></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">15797323</idno><idno type="pmc">7126974</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7126974</idno><idno type="RBID">PMC:7126974</idno><idno type="doi">10.1016/j.bios.2004.10.019</idno><date when="2004">2004</date><idno type="wicri:Area/Pmc/Corpus">000E58</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000E58</idno><idno type="wicri:Area/Pmc/Curation">000E58</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000E58</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Genosensor on gold films with enzymatic electrochemical detection of a SARS virus sequence<sup><xref ref-type="fn" rid="d32e1184">☆</xref></sup></title><author><name sortKey="Abad Valle, Patricia" sort="Abad Valle, Patricia" uniqKey="Abad Valle P" first="Patricia" last="Abad-Valle">Patricia Abad-Valle</name></author><author><name sortKey="Fernandez Abedul, M Teresa" sort="Fernandez Abedul, M Teresa" uniqKey="Fernandez Abedul M" first="M. Teresa" last="Fernández-Abedul">M. Teresa Fernández-Abedul</name></author><author><name sortKey="Costa Garcia, Agustin" sort="Costa Garcia, Agustin" uniqKey="Costa Garcia A" first="Agustín" last="Costa-García">Agustín Costa-García</name></author></analytic><series><title level="j">Biosensors & Bioelectronics</title><idno type="ISSN">0956-5663</idno><idno type="eISSN">1873-4235</idno><imprint><date when="2004">2004</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>A hybridisation-based genosensor was designed on a 100 nm sputtered gold film. This material worked as an immobilisation and transduction surface. A 30-mer sequence that encodes a short lysine-rich region, unique to SARS (severe acute respiratory syndrome) virus, was chosen as target. A complementary strand (probe), labelled with a thiol group at the 3′-end, was immobilised on the film. After blocking the surface, hybridisation with the biotin-conjugated SARS strand (at the 3′-end) took place. Interaction with alkaline phosphatase-labelled streptavidin permits amplified indirect electrochemical detection. The analytical signal is constituted by an electrochemical process of indigo carmine, the soluble product of the enzymatic hydrolysis of 3-indoxyl phosphate. The use of a sensitive electrochemical technique such as square wave voltammetry allowed a detection limit of 6 pM to be obtained for this DNA sequence, lower than any other found in the bibliography. The parameters affecting the methodology were studied, with special attention being placed on selectivity. Specificity was clearly enhanced when interaction time and stringency (in the form of formamide percentage) were increased. With 1 h of strand interaction and employing 50% of formamide in the hybridisation buffer, a 3-base mismatch strand was perfectly distinguished from the complementary.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Abad Villar, E M" uniqKey="Abad Villar E">E.M. Abad-Villar</name></author><author><name sortKey="Fernandez Abedul, M T" uniqKey="Fernandez Abedul M">M.T. Fernández-Abedul</name></author><author><name sortKey="Costa Garcia, A" uniqKey="Costa Garcia A">A. Costa-García</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Abad Villar, E M" uniqKey="Abad Villar E">E.M. Abad-Villar</name></author><author><name sortKey="Fernandez Abedul, M T" uniqKey="Fernandez Abedul M">M.T. Fernández-Abedul</name></author><author><name sortKey="Costa Garcia, A" uniqKey="Costa Garcia A">A. Costa-García</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Abad Villar, E M" uniqKey="Abad Villar E">E.M. Abad-Villar</name></author><author><name sortKey="Fernandez Abedul, M T" uniqKey="Fernandez Abedul M">M.T. Fernández-Abedul</name></author><author><name sortKey="Costa Garcia, A" uniqKey="Costa Garcia A">A. Costa-García</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Adams, R N" uniqKey="Adams R">R.N. Adams</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Aguilar, Z P" uniqKey="Aguilar Z">Z.P. Aguilar</name></author><author><name sortKey="Ritsch, I" uniqKey="Ritsch I">I. Ritsch</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Alfonta, L" uniqKey="Alfonta L">L. Alfonta</name></author><author><name sortKey="Singh, A K" uniqKey="Singh A">A.K. Singh</name></author><author><name sortKey="Willner, I" uniqKey="Willner I">I. Willner</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bain, C D" uniqKey="Bain C">C.D. Bain</name></author><author><name sortKey="Troughton, E B" uniqKey="Troughton E">E.B. Troughton</name></author><author><name sortKey="Tao, Y T" uniqKey="Tao Y">Y.-T. Tao</name></author><author><name sortKey="Evall, J" uniqKey="Evall J">J. Evall</name></author><author><name sortKey="Whitesides, G M" uniqKey="Whitesides G">G.M. Whitesides</name></author><author><name sortKey="Nuzzo, R G" uniqKey="Nuzzo R">R.G. Nuzzo</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bengoechea Lvarez, M J" uniqKey="Bengoechea Lvarez M">M.J. Bengoechea-Álvarez</name></author><author><name sortKey="Fernandez Abedul, M T" uniqKey="Fernandez Abedul M">M.T. Fernández-Abedul</name></author><author><name sortKey="Costa Garcia, A" uniqKey="Costa Garcia A">A. Costa-García</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Brett, A M O" uniqKey="Brett A">A.M.O. Brett</name></author><author><name sortKey="Matysik, F M" uniqKey="Matysik F">F.-M. Matysik</name></author><author><name sortKey="Vieira, M T" uniqKey="Vieira M">M.T. Vieira</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Campas, M" uniqKey="Campas M">M. Campas</name></author><author><name sortKey="Katakis, I" uniqKey="Katakis I">I. Katakis</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Carpini, G" uniqKey="Carpini G">G. Carpini</name></author><author><name sortKey="Lucarelli, F" uniqKey="Lucarelli F">F. Lucarelli</name></author><author><name sortKey="Marrazza, G" uniqKey="Marrazza G">G. Marrazza</name></author><author><name sortKey="Mascini, M" uniqKey="Mascini M">M. Mascini</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Casta O Lvarez, M" uniqKey="Casta O Lvarez M">M. Castaño-Álvarez</name></author><author><name sortKey="Fernandez Abedul, M T" uniqKey="Fernandez Abedul M">M.T. Fernández-Abedul</name></author><author><name sortKey="Costa Garcia, A" uniqKey="Costa Garcia A">A. Costa-García</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="De Los Santos Alvarez, P" uniqKey="De Los Santos Alvarez P">P. De-Los-Santos-Alvarez</name></author><author><name sortKey="Lobo Castanon, M J" uniqKey="Lobo Castanon M">M.J. Lobo-Castanon</name></author><author><name sortKey="Miranda Ordieres, A J" uniqKey="Miranda Ordieres A">A.J. Miranda-Ordieres</name></author><author><name sortKey="Tu N Blanco, P" uniqKey="Tu N Blanco P">P. Tuñón-Blanco</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Diaz Gonzalez, M" uniqKey="Diaz Gonzalez M">M. Díaz-González</name></author><author><name sortKey="Fernandez Sanchez, C" uniqKey="Fernandez Sanchez C">C. Fernández-Sánchez</name></author><author><name sortKey="Costa Garcia, A" uniqKey="Costa Garcia A">A. Costa-García</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Dominguez, E" uniqKey="Dominguez E">E. Domínguez</name></author><author><name sortKey="Rinc N, O" uniqKey="Rinc N O">O. Rincón</name></author><author><name sortKey="Narvaez, A" uniqKey="Narvaez A">A. Narváez</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Duan, C" uniqKey="Duan C">C. Duan</name></author><author><name sortKey="Meyerhoff, M E" uniqKey="Meyerhoff M">M.E. Meyerhoff</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Fernandez Sanchez, C" uniqKey="Fernandez Sanchez C">C. Fernández-Sánchez</name></author><author><name sortKey="Costa Garcia, A" uniqKey="Costa Garcia A">A. Costa-García</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Gherghi, I Ch" uniqKey="Gherghi I">I.Ch. Gherghi</name></author><author><name sortKey="Girousi, S Th" uniqKey="Girousi S">S.Th. Girousi</name></author><author><name sortKey="Voulgaropoulos, A" uniqKey="Voulgaropoulos A">A. Voulgaropoulos</name></author><author><name sortKey="Tzimou Tsitouridou, R" uniqKey="Tzimou Tsitouridou R">R. Tzimou-Tsitouridou</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Herne, T M" uniqKey="Herne T">T.M. Herne</name></author><author><name sortKey="Tarlov, M J" uniqKey="Tarlov M">M.J. Tarlov</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Jenkins, S H" uniqKey="Jenkins S">S.H. Jenkins</name></author><author><name sortKey="Heineman, W R" uniqKey="Heineman W">W.R. Heineman</name></author><author><name sortKey="Halsall, H B" uniqKey="Halsall H">H.B. Halsall</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Jordan, C E" uniqKey="Jordan C">C.E. Jordan</name></author><author><name sortKey="Frutos, A G" uniqKey="Frutos A">A.G. Frutos</name></author><author><name sortKey="Thiel, A J" uniqKey="Thiel A">A.J. Thiel</name></author><author><name sortKey="Corn, R M" uniqKey="Corn R">R.M. Corn</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Kelley, S O" uniqKey="Kelley S">S.O. Kelley</name></author><author><name sortKey="Boon, E M" uniqKey="Boon E">E.M. Boon</name></author><author><name sortKey="Barton, J K" uniqKey="Barton J">J.K. Barton</name></author><author><name sortKey="Jakson, N M" uniqKey="Jakson N">N.M. Jakson</name></author><author><name sortKey="Hill, M G" uniqKey="Hill M">M.G. Hill</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Levicky, R" uniqKey="Levicky R">R. Levicky</name></author><author><name sortKey="Herne, T M" uniqKey="Herne T">T.M. Herne</name></author><author><name sortKey="Tarlow, M J" uniqKey="Tarlow M">M.J. Tarlow</name></author><author><name sortKey="Satija, S K" uniqKey="Satija S">S.K. Satija</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lucarelli, F" uniqKey="Lucarelli F">F. Lucarelli</name></author><author><name sortKey="Marrazza, G" uniqKey="Marrazza G">G. Marrazza</name></author><author><name sortKey="Turner, A P F" uniqKey="Turner A">A.P.F. Turner</name></author><author><name sortKey="Marcini, M" uniqKey="Marcini M">M. Marcini</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Marra, M A" uniqKey="Marra M">M.A. Marra</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Matisyk, F M" uniqKey="Matisyk F">F.-M. Matisyk</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Meusel, M" uniqKey="Meusel M">M. Meusel</name></author><author><name sortKey="Renneberg, R" uniqKey="Renneberg R">R. Renneberg</name></author><author><name sortKey="Spener, F" uniqKey="Spener F">F. Spener</name></author><author><name sortKey="Schmitz, G" uniqKey="Schmitz G">G. Schmitz</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Minunni, M" uniqKey="Minunni M">M. Minunni</name></author><author><name sortKey="Tombelli, S" uniqKey="Tombelli S">S. Tombelli</name></author><author><name sortKey="Scielzi, R" uniqKey="Scielzi R">R. Scielzi</name></author><author><name sortKey="Mannelli, I" uniqKey="Mannelli I">I. Mannelli</name></author><author><name sortKey="Mascini, M" uniqKey="Mascini M">M. Mascini</name></author><author><name sortKey="Gaudiano, C" uniqKey="Gaudiano C">C. Gaudiano</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Nakamura, H" uniqKey="Nakamura H">H. Nakamura</name></author><author><name sortKey="Karube, I" uniqKey="Karube I">I. Karube</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Pan, W" uniqKey="Pan W">W. Pan</name></author><author><name sortKey="Durning, C J" uniqKey="Durning C">C.J. Durning</name></author><author><name sortKey="Turro, N J" uniqKey="Turro N">N.J. Turro</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Pividori, M I" uniqKey="Pividori M">M.I. Pividori</name></author><author><name sortKey="Merkoci, A" uniqKey="Merkoci A">A. Merkoçi</name></author><author><name sortKey="Alegret, S" uniqKey="Alegret S">S. Alegret</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Pividori, M I" uniqKey="Pividori M">M.I. Pividori</name></author><author><name sortKey="Merkoci" uniqKey="Merkoci">Merkoçi</name></author><author><name sortKey="Alegret, S" uniqKey="Alegret S">S. Alegret</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Steel, A B" uniqKey="Steel A">A.B. Steel</name></author><author><name sortKey="Herne, T M" uniqKey="Herne T">T.M. Herne</name></author><author><name sortKey="Tarlov, M J" uniqKey="Tarlov M">M.J. Tarlov</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Taft, B J" uniqKey="Taft B">B.J. Taft</name></author><author><name sortKey="O Eefe, M" uniqKey="O Eefe M">M. O’Keefe</name></author><author><name sortKey="Fourkas, J T" uniqKey="Fourkas J">J.T. Fourkas</name></author><author><name sortKey="Kelley, S O" uniqKey="Kelley S">S.O. Kelley</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Tie, F" uniqKey="Tie F">F. Tie</name></author><author><name sortKey="Pan, A" uniqKey="Pan A">A. Pan</name></author><author><name sortKey="Ru, B" uniqKey="Ru B">B. Ru</name></author><author><name sortKey="Wang, W" uniqKey="Wang W">W. Wang</name></author><author><name sortKey="Hu, Y" uniqKey="Hu Y">Y. Hu</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Wang, J" uniqKey="Wang J">J. Wang</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Wijayawardhana, C A" uniqKey="Wijayawardhana C">C.A. Wijayawardhana</name></author><author><name sortKey="Halsall, H B" uniqKey="Halsall H">H.B. Halsall</name></author><author><name sortKey="Heineman, W R" uniqKey="Heineman W">W.R. Heineman</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Wu, K" uniqKey="Wu K">K. Wu</name></author><author><name sortKey="Fei, J" uniqKey="Fei J">J. Fei</name></author><author><name sortKey="Bai, J" uniqKey="Bai J">J. Bai</name></author><author><name sortKey="Hu, S" uniqKey="Hu S">S. Hu</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Xu, D" uniqKey="Xu D">D. Xu</name></author><author><name sortKey="Huang, K" uniqKey="Huang K">K. Huang</name></author><author><name sortKey="Liu, Z" uniqKey="Liu Z">Z. Liu</name></author><author><name sortKey="Liu, Y" uniqKey="Liu Y">Y. Liu</name></author><author><name sortKey="Ma, L" uniqKey="Ma L">L. Ma</name></author></analytic></biblStruct><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Zhu, N" uniqKey="Zhu N">N. Zhu</name></author><author><name sortKey="Cai, H" uniqKey="Cai H">H. Cai</name></author><author><name sortKey="He, P" uniqKey="He P">P. He</name></author><author><name sortKey="Fang, Y" uniqKey="Fang Y">Y. Fang</name></author></analytic></biblStruct></listBibl></div1></back></TEI><pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Biosens Bioelectron</journal-id><journal-id journal-id-type="iso-abbrev">Biosens Bioelectron</journal-id><journal-title-group><journal-title>Biosensors & Bioelectronics</journal-title></journal-title-group><issn pub-type="ppub">0956-5663</issn><issn pub-type="epub">1873-4235</issn><publisher><publisher-name>Elsevier B.V.</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">15797323</article-id><article-id pub-id-type="pmc">7126974</article-id><article-id pub-id-type="publisher-id">S0956-5663(04)00512-3</article-id><article-id pub-id-type="doi">10.1016/j.bios.2004.10.019</article-id><article-categories><subj-group subj-group-type="heading"><subject>Article</subject></subj-group></article-categories><title-group><article-title>Genosensor on gold films with enzymatic electrochemical detection of a SARS virus sequence<sup><xref ref-type="fn" rid="d32e1184">☆</xref></sup></article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Abad-Valle</surname><given-names>Patricia</given-names></name></contrib><contrib contrib-type="author"><name><surname>Fernández-Abedul</surname><given-names>M. Teresa</given-names></name></contrib><contrib contrib-type="author"><name><surname>Costa-García</surname><given-names>Agustín</given-names></name><email>costa@fq.uniovi.es</email><xref rid="cor1" ref-type="corresp">*</xref></contrib></contrib-group><aff>Departamento de Química Física y Analítica, Universidad de Oviedo, Asturias, 33006 Oviedo, Spain</aff><author-notes><corresp id="cor1"><label>*</label>Corresponding author. Tel.: +34 985103488; fax: +34 985103125. <email>costa@fq.uniovi.es</email></corresp></author-notes><pub-date pub-type="pmc-release"><day>13</day><month>12</month><year>2004</year></pub-date><pmc-comment> PMC Release delay is 0 months and 0 days and was based on .</pmc-comment>
<pub-date pub-type="ppub"><day>15</day><month>5</month><year>2005</year></pub-date><pub-date pub-type="epub"><day>13</day><month>12</month><year>2004</year></pub-date><volume>20</volume><issue>11</issue><fpage>2251</fpage><lpage>2260</lpage><history><date date-type="received"><day>29</day><month>7</month><year>2004</year></date><date date-type="rev-recd"><day>21</day><month>10</month><year>2004</year></date><date date-type="accepted"><day>22</day><month>10</month><year>2004</year></date></history><permissions><copyright-statement>Copyright © 2004 Elsevier B.V. All rights reserved.</copyright-statement><copyright-year>2004</copyright-year><copyright-holder>Elsevier B.V.</copyright-holder><license><license-p>Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.</license-p></license></permissions><abstract><p>A hybridisation-based genosensor was designed on a 100 nm sputtered gold film. This material worked as an immobilisation and transduction surface. A 30-mer sequence that encodes a short lysine-rich region, unique to SARS (severe acute respiratory syndrome) virus, was chosen as target. A complementary strand (probe), labelled with a thiol group at the 3′-end, was immobilised on the film. After blocking the surface, hybridisation with the biotin-conjugated SARS strand (at the 3′-end) took place. Interaction with alkaline phosphatase-labelled streptavidin permits amplified indirect electrochemical detection. The analytical signal is constituted by an electrochemical process of indigo carmine, the soluble product of the enzymatic hydrolysis of 3-indoxyl phosphate. The use of a sensitive electrochemical technique such as square wave voltammetry allowed a detection limit of 6 pM to be obtained for this DNA sequence, lower than any other found in the bibliography. The parameters affecting the methodology were studied, with special attention being placed on selectivity. Specificity was clearly enhanced when interaction time and stringency (in the form of formamide percentage) were increased. With 1 h of strand interaction and employing 50% of formamide in the hybridisation buffer, a 3-base mismatch strand was perfectly distinguished from the complementary.</p></abstract><kwd-group><title>Keywords</title><kwd>DNA</kwd><kwd>Hybridisation assay</kwd><kwd>Genosensor</kwd><kwd>Enzyme assay</kwd><kwd>Gold film</kwd><kwd>Electrochemical detection (ED)</kwd><kwd>SARS</kwd></kwd-group></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/SrasV1/Data/Pmc/Curation
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000E58 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Pmc/Curation/biblio.hfd -nk 000E58 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= SrasV1
|flux= Pmc
|étape= Curation
|type= RBID
|clé= PMC:7126974
|texte= Genosensor on gold films with enzymatic electrochemical detection of a SARS virus sequence☆
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Pmc/Curation/RBID.i -Sk "pubmed:15797323" \
| HfdSelect -Kh $EXPLOR_AREA/Data/Pmc/Curation/biblio.hfd \
| NlmPubMed2Wicri -a SrasV1
| This area was generated with Dilib version V0.6.33. |  |