A Real-Time PCR Assay for Bat SARS-Like Coronavirus Detection and Its Application to Italian Greater Horseshoe Bat Faecal Sample Surveys
Identifieur interne : 001475 ( Pmc/Curation ); précédent : 001474; suivant : 001476A Real-Time PCR Assay for Bat SARS-Like Coronavirus Detection and Its Application to Italian Greater Horseshoe Bat Faecal Sample Surveys
Auteurs : Andrea Balboni [Italie] ; Laura Gallina [Italie] ; Alessandra Palladini [Italie] ; Santino Prosperi [Italie] ; Mara Battilani [Italie]Source :
- The Scientific World Journal [ 2356-6140 ] ; 2011.
Abstract
Bats are source of coronaviruses closely related to the severe acute respiratory syndrome (SARS) virus. Numerous studies have been carried out to identify new bat viruses related to SARS-coronavirus (bat-SARS-like CoVs) using a reverse-transcribed-polymerase chain reaction assay. However, a qualitative PCR could underestimate the prevalence of infection, affecting the epidemiological evaluation of bats in viral ecology. In this work an SYBR Green-real time PCR assay was developed for diagnosing infection with SARS-related coronaviruses from bat guano and was applied as screening tool in a survey carried out on 45 greater horseshoe bats (
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DOI: 10.1100/2012/989514
PubMed: 22654650
PubMed Central: 3353321
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">A Real-Time PCR Assay for Bat SARS-Like Coronavirus Detection and
Its Application to Italian Greater Horseshoe Bat Faecal Sample Surveys</title><author><name sortKey="Balboni, Andrea" sort="Balboni, Andrea" uniqKey="Balboni A" first="Andrea" last="Balboni">Andrea Balboni</name><affiliation wicri:level="1"><nlm:aff id="I1">Dipartimento di Scienze Mediche Veterinarie, Alma Mater Studiorum-Università di Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Dipartimento di Scienze Mediche Veterinarie, Alma Mater Studiorum-Università di Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia</wicri:regionArea></affiliation></author><author><name sortKey="Gallina, Laura" sort="Gallina, Laura" uniqKey="Gallina L" first="Laura" last="Gallina">Laura Gallina</name><affiliation wicri:level="1"><nlm:aff id="I1">Dipartimento di Scienze Mediche Veterinarie, Alma Mater Studiorum-Università di Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Dipartimento di Scienze Mediche Veterinarie, Alma Mater Studiorum-Università di Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia</wicri:regionArea></affiliation></author><author><name sortKey="Palladini, Alessandra" sort="Palladini, Alessandra" uniqKey="Palladini A" first="Alessandra" last="Palladini">Alessandra Palladini</name><affiliation wicri:level="1"><nlm:aff id="I2">Dipartimento di Biologia Animale, Università degli Studi di Pavia, Via Taramelli 24, 27100 Pavia, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Dipartimento di Biologia Animale, Università degli Studi di Pavia, Via Taramelli 24, 27100 Pavia</wicri:regionArea></affiliation></author><author><name sortKey="Prosperi, Santino" sort="Prosperi, Santino" uniqKey="Prosperi S" first="Santino" last="Prosperi">Santino Prosperi</name><affiliation wicri:level="1"><nlm:aff id="I1">Dipartimento di Scienze Mediche Veterinarie, Alma Mater Studiorum-Università di Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Dipartimento di Scienze Mediche Veterinarie, Alma Mater Studiorum-Università di Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia</wicri:regionArea></affiliation></author><author><name sortKey="Battilani, Mara" sort="Battilani, Mara" uniqKey="Battilani M" first="Mara" last="Battilani">Mara Battilani</name><affiliation wicri:level="1"><nlm:aff id="I1">Dipartimento di Scienze Mediche Veterinarie, Alma Mater Studiorum-Università di Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Dipartimento di Scienze Mediche Veterinarie, Alma Mater Studiorum-Università di Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia</wicri:regionArea></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">22654650</idno><idno type="pmc">3353321</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3353321</idno><idno type="RBID">PMC:3353321</idno><idno type="doi">10.1100/2012/989514</idno><date when="2011">2011</date><idno type="wicri:Area/Pmc/Corpus">001475</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">001475</idno><idno type="wicri:Area/Pmc/Curation">001475</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">001475</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">A Real-Time PCR Assay for Bat SARS-Like Coronavirus Detection and
Its Application to Italian Greater Horseshoe Bat Faecal Sample Surveys</title><author><name sortKey="Balboni, Andrea" sort="Balboni, Andrea" uniqKey="Balboni A" first="Andrea" last="Balboni">Andrea Balboni</name><affiliation wicri:level="1"><nlm:aff id="I1">Dipartimento di Scienze Mediche Veterinarie, Alma Mater Studiorum-Università di Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Dipartimento di Scienze Mediche Veterinarie, Alma Mater Studiorum-Università di Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia</wicri:regionArea></affiliation></author><author><name sortKey="Gallina, Laura" sort="Gallina, Laura" uniqKey="Gallina L" first="Laura" last="Gallina">Laura Gallina</name><affiliation wicri:level="1"><nlm:aff id="I1">Dipartimento di Scienze Mediche Veterinarie, Alma Mater Studiorum-Università di Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Dipartimento di Scienze Mediche Veterinarie, Alma Mater Studiorum-Università di Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia</wicri:regionArea></affiliation></author><author><name sortKey="Palladini, Alessandra" sort="Palladini, Alessandra" uniqKey="Palladini A" first="Alessandra" last="Palladini">Alessandra Palladini</name><affiliation wicri:level="1"><nlm:aff id="I2">Dipartimento di Biologia Animale, Università degli Studi di Pavia, Via Taramelli 24, 27100 Pavia, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Dipartimento di Biologia Animale, Università degli Studi di Pavia, Via Taramelli 24, 27100 Pavia</wicri:regionArea></affiliation></author><author><name sortKey="Prosperi, Santino" sort="Prosperi, Santino" uniqKey="Prosperi S" first="Santino" last="Prosperi">Santino Prosperi</name><affiliation wicri:level="1"><nlm:aff id="I1">Dipartimento di Scienze Mediche Veterinarie, Alma Mater Studiorum-Università di Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Dipartimento di Scienze Mediche Veterinarie, Alma Mater Studiorum-Università di Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia</wicri:regionArea></affiliation></author><author><name sortKey="Battilani, Mara" sort="Battilani, Mara" uniqKey="Battilani M" first="Mara" last="Battilani">Mara Battilani</name><affiliation wicri:level="1"><nlm:aff id="I1">Dipartimento di Scienze Mediche Veterinarie, Alma Mater Studiorum-Università di Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Dipartimento di Scienze Mediche Veterinarie, Alma Mater Studiorum-Università di Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia</wicri:regionArea></affiliation></author></analytic><series><title level="j">The Scientific World Journal</title><idno type="ISSN">2356-6140</idno><idno type="eISSN">1537-744X</idno><imprint><date when="2011">2011</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Bats are source of coronaviruses closely related to the severe acute respiratory syndrome (SARS) virus. Numerous studies have been carried out to identify new bat viruses related to SARS-coronavirus (bat-SARS-like CoVs) using a reverse-transcribed-polymerase chain reaction assay. However, a qualitative PCR could underestimate the prevalence of infection, affecting the epidemiological evaluation of bats in viral ecology. In this work an SYBR Green-real time PCR assay was developed for diagnosing infection with SARS-related coronaviruses from bat guano and was applied as screening tool in a survey carried out on 45 greater horseshoe bats (<italic>Rhinolophus ferrumequinum</italic>) sampled in Italy in 2009. The assay showed high sensitivity and reproducibility. Its application on bats screening resulted in a prevalence of 42%. This method could be suitable as screening tool in epidemiological surveys about the presence of bat-SARS-like CoVs, consequently to obtain a more realistic scenario of the viral prevalence in the population.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Peiris, Jsm" uniqKey="Peiris J">JSM Peiris</name></author><author><name sortKey="Guan, Y" uniqKey="Guan Y">Y Guan</name></author><author><name sortKey="Yuen, Ky" uniqKey="Yuen K">KY Yuen</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Rota, Pa" uniqKey="Rota P">PA Rota</name></author><author><name sortKey="Oberste, Ms" uniqKey="Oberste M">MS Oberste</name></author><author><name sortKey="Monroe, Ss" uniqKey="Monroe S">SS Monroe</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Shi, Z" uniqKey="Shi Z">Z Shi</name></author><author><name sortKey="Hu, Z" uniqKey="Hu Z">Z 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<front><journal-meta><journal-id journal-id-type="nlm-ta">ScientificWorldJournal</journal-id><journal-id journal-id-type="iso-abbrev">ScientificWorldJournal</journal-id><journal-id journal-id-type="publisher-id">TSWJ</journal-id><journal-title-group><journal-title>The Scientific World Journal</journal-title></journal-title-group><issn pub-type="ppub">2356-6140</issn><issn pub-type="epub">1537-744X</issn><publisher><publisher-name>The Scientific World Journal</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">22654650</article-id><article-id pub-id-type="pmc">3353321</article-id><article-id pub-id-type="doi">10.1100/2012/989514</article-id><article-categories><subj-group subj-group-type="heading"><subject>Research Article</subject></subj-group></article-categories><title-group><article-title>A Real-Time PCR Assay for Bat SARS-Like Coronavirus Detection and
Its Application to Italian Greater Horseshoe Bat Faecal Sample Surveys</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Balboni</surname><given-names>Andrea</given-names></name><xref ref-type="aff" rid="I1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Gallina</surname><given-names>Laura</given-names></name><xref ref-type="aff" rid="I1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Palladini</surname><given-names>Alessandra</given-names></name><xref ref-type="aff" rid="I2"><sup>2</sup></xref></contrib><contrib contrib-type="author"><name><surname>Prosperi</surname><given-names>Santino</given-names></name><xref ref-type="aff" rid="I1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Battilani</surname><given-names>Mara</given-names></name><xref ref-type="aff" rid="I1"><sup>1</sup></xref><xref ref-type="corresp" rid="cor1">*</xref></contrib></contrib-group><aff id="I1"><sup>1</sup>Dipartimento di Scienze Mediche Veterinarie, Alma Mater Studiorum-Università di Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia, Italy</aff><aff id="I2"><sup>2</sup>Dipartimento di Biologia Animale, Università degli Studi di Pavia, Via Taramelli 24, 27100 Pavia, Italy</aff><author-notes><corresp id="cor1">*Mara Battilani: <email>mara.battilani@unibo.it</email></corresp><fn fn-type="other"><p>Academic Editors: R. Marquet, M. McCrae, and S. Y. Morozov</p></fn></author-notes><pub-date pub-type="collection"><year>2012</year></pub-date><pub-date pub-type="epub"><day>22</day><month>11</month><year>2011</year></pub-date><volume>2012</volume><elocation-id>989514</elocation-id><history><date date-type="received"><day>5</day><month>10</month><year>2011</year></date><date date-type="accepted"><day>16</day><month>11</month><year>2011</year></date></history><permissions><copyright-statement>Copyright © 2012 Andrea Balboni et al.</copyright-statement><copyright-year>2012</copyright-year><license xlink:href="https://creativecommons.org/licenses/by/3.0/"><license-p>This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</license-p></license></permissions><abstract><p>Bats are source of coronaviruses closely related to the severe acute respiratory syndrome (SARS) virus. Numerous studies have been carried out to identify new bat viruses related to SARS-coronavirus (bat-SARS-like CoVs) using a reverse-transcribed-polymerase chain reaction assay. However, a qualitative PCR could underestimate the prevalence of infection, affecting the epidemiological evaluation of bats in viral ecology. In this work an SYBR Green-real time PCR assay was developed for diagnosing infection with SARS-related coronaviruses from bat guano and was applied as screening tool in a survey carried out on 45 greater horseshoe bats (<italic>Rhinolophus ferrumequinum</italic>) sampled in Italy in 2009. The assay showed high sensitivity and reproducibility. Its application on bats screening resulted in a prevalence of 42%. This method could be suitable as screening tool in epidemiological surveys about the presence of bat-SARS-like CoVs, consequently to obtain a more realistic scenario of the viral prevalence in the population.</p></abstract></article-meta></front><floats-group><fig id="fig1" position="float"><label>Figure 1</label><caption><p>Distribution of bat sampling in Italy. Sites of bat sampling from August to November 2009. For correspondence between letters and places, see <xref ref-type="table" rid="tab1">Table 1</xref>.</p></caption><graphic xlink:href="TSWJ2012-989514.001"></graphic></fig><fig id="fig2" position="float"><label>Figure 2</label><caption><p>Multiple sequence alignment of primer binding sites. The alignments include 10 SARS-related coronavirus reference strains: SBRp3 (Bat SARS CoV Rp3/2004, DQ071615, identified from <italic>Rhinolophus pearsoni</italic>), SBRs (SARS coronavirus Rs_672/2006, FJ588686, identified from <italic>Rhinolophus sinicus</italic>), SHTor2 (SARS coronavirus Tor2, AY274119, identified from Human), SC (Civet SARS CoV SZ16/2003, AY304488, identified from<italic> Paguma larvata</italic>), SB (SARS coronavirus isolate CFB/SZ/94/03, AY545919, identified from<italic> Melogale moschata</italic>), SBRf1 (Bat SARS CoV Rf1/2004, DQ412042, identified from <italic>Rhinolophus ferrumequinum</italic>), SBRm1 (Bat SARS CoV Rm1/2004, DQ412043, identified from <italic>Rhinolophus macrotis</italic>), SBHKU3-1 (Bat coronavirus HKU3, DQ022305, identified from <italic>Rhinolophus sinicus</italic>), SB273 (Bat CoV 273/2005, DQ648856, identified from <italic>Rhinolophus ferrumequinum</italic>), and SB279 (Bat CoV 279/2005, DQ648857, identified from <italic>Rhinolophus macrotis</italic>).</p></caption><graphic xlink:href="TSWJ2012-989514.002"></graphic></fig><fig id="fig3" position="float"><label>Figure 3</label><caption><p>Real-time PCR reaction products checked on agarose gel stained with ethidium bromide in standard tris-acetate-EDTA (TAE) buffer: serial 10-fold dilutions of recombinant plasmid. Specific bands of approximately 168 bp were visualised for all replicates of recombinant plasmid dilutions, except the concentration of 1 × 10<sup>−1</sup> copies/<italic>μ</italic>L. For recombinant plasmid dilution with a concentration of 1 × 10<sup>−1</sup> copies/<italic>μ</italic>L only one specific amplicon to three replicates was visualised. MK: MassRuler Low-Range DNA Ladder (Fermentas, Burlington, Ontario, Canada). a, b, c, d, and e: recombinant plasmid dilutions with concentrations of 1 × 10<sup>3</sup>, 1 × 10<sup>2</sup>, 1 × 10<sup>1</sup>, 1 × 10<sup>0</sup>, and 1 × 10<sup>−1</sup> copies/<italic>μ</italic>L, respectively. W: no template control (water).</p></caption><graphic xlink:href="TSWJ2012-989514.003"></graphic></fig><fig id="fig4" position="float"><label>Figure 4</label><caption><p>Melting curve analysis of standard plasmid dilutions and sample 893/09-11. In gray: signals obtained from the standard plasmid dilutions; in black: signals obtained from the sample 893.09-11; derivative −<italic>dF</italic>/<italic>dT</italic> where <italic>F</italic> is fluorescence and <italic>T</italic> is time; °C: temperature (centigrade).</p></caption><graphic xlink:href="TSWJ2012-989514.004"></graphic></fig><fig id="fig5" position="float"><label>Figure 5</label><caption><p>Real time-PCR reaction products checked on agarose gel stained with ethidium bromide in standard tris-acetate-EDTA (TAE) buffer: positive samples detected by developed real-time PCR. Specific bands of approximately 168 bp were visualised for all replicates of the detected positive samples. In this figure is represented the 2% (w/v) agarose gel electrophoresis of five of the 11 positive samples detected in bats belonging to sampling area A (San Cesario sul Panaro, MO). Pl: amplicon of the recombinant plasmid with 1 × 10<sup>4</sup> copies/<italic>μ</italic>L; 1, 2, 3, 4, and 5: amplicons of the five positive samples belonging to sampling area A, each with three repetitions.</p></caption><graphic xlink:href="TSWJ2012-989514.005"></graphic></fig><table-wrap id="tab1" position="float"><label>Table 1</label><caption><p>Bats tested for coronavirus infection using SYBR Green real-time PCR assay.</p></caption><table frame="hsides" rules="groups"><thead><tr><th align="left" rowspan="1" colspan="1"></th><th align="left" rowspan="1" colspan="1">Location</th><th align="center" rowspan="1" colspan="1">Date</th><th align="center" rowspan="1" colspan="1">No. of bats</th><th align="center" rowspan="1" colspan="1">Positives</th></tr></thead><tbody><tr><td align="left" rowspan="1" colspan="1">A</td><td align="left" rowspan="1" colspan="1">San Cesario sul Panaro, MO</td><td align="center" rowspan="1" colspan="1">16/08/2009</td><td align="center" rowspan="1" colspan="1">11</td><td align="center" rowspan="1" colspan="1">10</td></tr><tr><td align="left" rowspan="1" colspan="1">B</td><td align="left" rowspan="1" colspan="1">Sigillo, PG</td><td align="center" rowspan="1" colspan="1">18/09/2009</td><td align="center" rowspan="1" colspan="1">1</td><td align="center" rowspan="1" colspan="1">0</td></tr><tr><td align="left" rowspan="1" colspan="1">C</td><td align="left" rowspan="1" colspan="1">Monte Croara, S. Lazzaro, BO</td><td align="center" rowspan="1" colspan="1">05/10/2009</td><td align="center" rowspan="1" colspan="1">2</td><td align="center" rowspan="1" colspan="1">0</td></tr><tr><td align="left" rowspan="1" colspan="1">D</td><td align="left" rowspan="1" colspan="1">Piobesi d'Alba, CN</td><td align="center" rowspan="1" colspan="1">11/10/2009</td><td align="center" rowspan="1" colspan="1">10</td><td align="center" rowspan="1" colspan="1">0</td></tr><tr><td align="left" rowspan="1" colspan="1">E</td><td align="left" rowspan="1" colspan="1">Rossana, CN</td><td align="center" rowspan="1" colspan="1">11/10/2009</td><td align="center" rowspan="1" colspan="1">12</td><td align="center" rowspan="1" colspan="1">3</td></tr><tr><td align="left" rowspan="1" colspan="1">F</td><td align="left" rowspan="1" colspan="1">Giovo, SV</td><td align="center" rowspan="1" colspan="1">19/10/2009</td><td align="center" rowspan="1" colspan="1">3</td><td align="center" rowspan="1" colspan="1">3</td></tr><tr><td align="left" rowspan="1" colspan="1">G</td><td align="left" rowspan="1" colspan="1">Val di Trebbia, PC</td><td align="center" rowspan="1" colspan="1">20/10/2009</td><td align="center" rowspan="1" colspan="1">3</td><td align="center" rowspan="1" colspan="1">1</td></tr><tr><td align="left" rowspan="1" colspan="1">H</td><td align="left" rowspan="1" colspan="1">Castell'arquato, PC</td><td align="center" rowspan="1" colspan="1">04/11/2009</td><td align="center" rowspan="1" colspan="1">3</td><td align="center" rowspan="1" colspan="1">2</td></tr><tr><td align="left" colspan="5" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"></td><td align="left" rowspan="1" colspan="1">Total</td><td align="center" rowspan="1" colspan="1"></td><td align="center" rowspan="1" colspan="1">45</td><td align="center" rowspan="1" colspan="1">19</td></tr></tbody></table></table-wrap><table-wrap id="tab2" position="float"><label>Table 2</label><caption><p>Intra- and interassay variability of the SYBR green real-time PCR method.</p></caption><table frame="hsides" rules="groups"><thead><tr><th align="left" colspan="5" rowspan="1">Intra- and interassay variability</th></tr></thead><tbody><tr><td align="left" rowspan="1" colspan="1">Samples</td><td align="center" rowspan="1" colspan="1">Replicate (and assay) numbers</td><td align="center" rowspan="1" colspan="1">Mean (SD)</td><td align="center" rowspan="1" colspan="1">log<sub>10</sub> mean (SD)</td><td align="center" rowspan="1" colspan="1">log<sub>10</sub> CV%</td></tr><tr><td align="left" colspan="5" rowspan="1"><hr></hr></td></tr><tr><td align="left" colspan="5" rowspan="1">Intraassay variability</td></tr><tr><td align="left" rowspan="1" colspan="1">A1</td><td align="center" rowspan="1" colspan="1">3 (1)</td><td align="center" rowspan="1" colspan="1">1,86<italic>E</italic> + 06 (2,35<italic>E</italic> + 05)</td><td align="center" rowspan="1" colspan="1">6,27 (0,05)</td><td align="center" rowspan="1" colspan="1">0,84</td></tr><tr><td align="left" rowspan="1" colspan="1">A2</td><td align="center" rowspan="1" colspan="1">3 (1)</td><td align="center" rowspan="1" colspan="1">2,03<italic>E</italic> + 05 (1,77<italic>E</italic> + 04)</td><td align="center" rowspan="1" colspan="1">5,31 (0,04)</td><td align="center" rowspan="1" colspan="1">0,71</td></tr><tr><td align="left" rowspan="1" colspan="1">A3</td><td align="center" rowspan="1" colspan="1">3 (1)</td><td align="center" rowspan="1" colspan="1">2,00<italic>E</italic> + 04 (8,96<italic>E</italic> + 02)</td><td align="center" rowspan="1" colspan="1">4,30 (0,02)</td><td align="center" rowspan="1" colspan="1">0,45</td></tr><tr><td align="left" rowspan="1" colspan="1">A4</td><td align="center" rowspan="1" colspan="1">3 (1)</td><td align="center" rowspan="1" colspan="1">2,26<italic>E</italic> + 03 (2,53<italic>E</italic> + 02)</td><td align="center" rowspan="1" colspan="1">3,35 (0,05)</td><td align="center" rowspan="1" colspan="1">1,41</td></tr><tr><td align="left" rowspan="1" colspan="1">A5</td><td align="center" rowspan="1" colspan="1">3 (1)</td><td align="center" rowspan="1" colspan="1">2,09<italic>E</italic> + 02 (8,22)</td><td align="center" rowspan="1" colspan="1">2,32 (0,02)</td><td align="center" rowspan="1" colspan="1">0,74</td></tr><tr><td align="left" rowspan="1" colspan="1">A6</td><td align="center" rowspan="1" colspan="1">3 (1)</td><td align="center" rowspan="1" colspan="1">1,98<italic>E</italic> + 01 (5,29)</td><td align="center" rowspan="1" colspan="1">1,28 (0,13)</td><td align="center" rowspan="1" colspan="1">9,99</td></tr><tr><td align="left" rowspan="1" colspan="1">A7</td><td align="center" rowspan="1" colspan="1">3 (1)</td><td align="center" rowspan="1" colspan="1">2,09<italic>E</italic> + 00 (0,44)</td><td align="center" rowspan="1" colspan="1">0,31 (0,09)</td><td align="center" rowspan="1" colspan="1">28,58</td></tr><tr><td align="left" rowspan="1" colspan="1">B1</td><td align="center" rowspan="1" colspan="1">3 (1)</td><td align="center" rowspan="1" colspan="1">5,79<italic>E</italic> + 02 (4,65<italic>E</italic> + 01)</td><td align="center" rowspan="1" colspan="1">2,76 (0,03)</td><td align="center" rowspan="1" colspan="1">1,27</td></tr><tr><td align="left" rowspan="1" colspan="1">B2</td><td align="center" rowspan="1" colspan="1">3 (1)</td><td align="center" rowspan="1" colspan="1">2,00<italic>E</italic> + 00 (0,10)</td><td align="center" rowspan="1" colspan="1">0,30 (0,02)</td><td align="center" rowspan="1" colspan="1">7,56</td></tr><tr><td align="left" rowspan="1" colspan="1">B3</td><td align="center" rowspan="1" colspan="1">3 (1)</td><td align="center" rowspan="1" colspan="1">4,01<italic>E</italic> + 00 (0,74)</td><td align="center" rowspan="1" colspan="1">0,60 (0,08)</td><td align="center" rowspan="1" colspan="1">13,68</td></tr><tr><td align="left" colspan="5" rowspan="1"><hr></hr></td></tr><tr><td align="left" colspan="5" rowspan="1">Interassay variability</td></tr><tr><td align="left" rowspan="1" colspan="1">B1</td><td align="center" rowspan="1" colspan="1">3 (3)</td><td align="center" rowspan="1" colspan="1">6,08<italic>E</italic> + 02 (5,58<italic>E</italic> + 01)</td><td align="center" rowspan="1" colspan="1">2,78 (0,04)</td><td align="center" rowspan="1" colspan="1">1,38</td></tr><tr><td align="left" rowspan="1" colspan="1">B2</td><td align="center" rowspan="1" colspan="1">3 (3)</td><td align="center" rowspan="1" colspan="1">2,06<italic>E</italic> + 00 (0,34)</td><td align="center" rowspan="1" colspan="1">0,29 (0,07)</td><td align="center" rowspan="1" colspan="1">23,97</td></tr><tr><td align="left" rowspan="1" colspan="1">B3</td><td align="center" rowspan="1" colspan="1">3 (3)</td><td align="center" rowspan="1" colspan="1">3,11<italic>E</italic> + 00 (0,82)</td><td align="center" rowspan="1" colspan="1">0,46 (0,13)</td><td align="center" rowspan="1" colspan="1">28,79</td></tr></tbody></table><table-wrap-foot><fn><p>Successive 10-fold dilutions of recombinant plasmid: A1 (2 × 10<sup>6</sup>), A2 (2 × 10<sup>5</sup>), A3 (2 × 10<sup>4</sup>), A4 (2 × 10<sup>3</sup>), A5 (2 × 10<sup>2</sup>), A6 (2 × 10<sup>1</sup>), and A7 (2 × 10<sup>0</sup>).</p></fn><fn><p>Three bat samples with different viral concentrations: B1 (10<sup>2</sup>), B2 and B3 (10<sup>0</sup>). </p></fn><fn><p>SD: standard deviation.</p></fn><fn><p>CV: coefficient of variation.</p></fn></table-wrap-foot></table-wrap></floats-group></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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