Specific detection of D- and M-isolates of plum pox virus by immunoenzymatic determination of PCR products
Identifieur interne : 003C41 ( Main/Exploration ); précédent : 003C40; suivant : 003C42Specific detection of D- and M-isolates of plum pox virus by immunoenzymatic determination of PCR products
Auteurs : C. Poggi Pollini [Italie] ; L. Giunchedi [Italie] ; R. Bissani [Italie]Source :
- Journal of Virological Methods [ 0166-0934 ] ; 1997.
English descriptors
- Teeft :
- Absorption values, Acta hortic, Antisense primer, Apricot, Apricot orchards, Apricot trees, Assay, Boehringer mannheim, Candresse, Clear discrimination, Detection limit, Elisa reader, Enzyme immunoassay, Epidemiological conditions, Eppo bull, Extraction protocol, Hazardous chemicals, Healthy samples, Human medicine, Hybridisation, Hybridisation assay, Immunoenzymatic determination, Incubation, Incubation buffer, Incubation temperature, Incubation time, Leaf samples, Mannheim, Negative results, Peach, Peach trees, Plant pathology, Plum, Plum trees, Poggi, Poggi pollini, Pollini, Polymerase chain reaction, Present study, Primer, Probe, Probe concentration, Rapid detection, Reaction mixture, Rflp, Rflp analysis, Spectrophotometric readings, Sterile water, Stone fruit trees, Subsequent rflp analysis, Sugar beet roots, Variable region, Virological, Virological methods, Woody plants.
Abstract
Abstract: Molecular techniques based on the polymerase chain reaction (PCR) can provide rapid and sensitive diagnosis of plum pox virus (PPV), the causal agent of the devastating `sharka' disease of stone fruit trees. The present study compared routine polymerase chain reaction (PCR) procedures against a new system, PCR-ELISA (Boehringer Mannheim), which enables immunoenzymatic detection of PCR products. The results show that this hybridisation system ensures fast and more sensitive detection of PPV associated with stone fruit trees and herbaceous hosts. Strain-specific capture probes were also designed to identify the two major PPV isolates, D and M, without subsequent restriction fragment length polymorphism analysis of the PCR products. Optimisation of all parameters involved in the PCR-ELISA procedure are discussed and its advantages reported.
Url:
DOI: 10.1016/S0166-0934(97)00085-2
Affiliations:
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Poggi Pollini</name><affiliation wicri:level="1"><country xml:lang="fr">Italie</country><wicri:regionArea>Istituto di Patologia Vegetale, via Filippo Re 8, 40126 Bologna</wicri:regionArea><wicri:noRegion>40126 Bologna</wicri:noRegion></affiliation></author><author><name sortKey="Giunchedi, L" sort="Giunchedi, L" uniqKey="Giunchedi L" first="L" last="Giunchedi">L. Giunchedi</name><affiliation wicri:level="1"><country xml:lang="fr">Italie</country><wicri:regionArea>Istituto di Patologia Vegetale, via Filippo Re 8, 40126 Bologna</wicri:regionArea><wicri:noRegion>40126 Bologna</wicri:noRegion></affiliation></author><author><name sortKey="Bissani, R" sort="Bissani, R" uniqKey="Bissani R" first="R" last="Bissani">R. Bissani</name><affiliation wicri:level="1"><country xml:lang="fr">Italie</country><wicri:regionArea>Istituto di Patologia Vegetale, via Filippo Re 8, 40126 Bologna</wicri:regionArea><wicri:noRegion>40126 Bologna</wicri:noRegion></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="1997">1997</date><biblScope unit="volume">67</biblScope><biblScope unit="issue">2</biblScope><biblScope unit="page" from="127">127</biblScope><biblScope unit="page" to="133">133</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>Absorption values</term><term>Acta hortic</term><term>Antisense primer</term><term>Apricot</term><term>Apricot orchards</term><term>Apricot trees</term><term>Assay</term><term>Boehringer mannheim</term><term>Candresse</term><term>Clear discrimination</term><term>Detection limit</term><term>Elisa reader</term><term>Enzyme immunoassay</term><term>Epidemiological conditions</term><term>Eppo bull</term><term>Extraction protocol</term><term>Hazardous chemicals</term><term>Healthy samples</term><term>Human medicine</term><term>Hybridisation</term><term>Hybridisation assay</term><term>Immunoenzymatic determination</term><term>Incubation</term><term>Incubation buffer</term><term>Incubation temperature</term><term>Incubation time</term><term>Leaf samples</term><term>Mannheim</term><term>Negative results</term><term>Peach</term><term>Peach trees</term><term>Plant pathology</term><term>Plum</term><term>Plum trees</term><term>Poggi</term><term>Poggi pollini</term><term>Pollini</term><term>Polymerase chain reaction</term><term>Present study</term><term>Primer</term><term>Probe</term><term>Probe concentration</term><term>Rapid detection</term><term>Reaction mixture</term><term>Rflp</term><term>Rflp analysis</term><term>Spectrophotometric readings</term><term>Sterile water</term><term>Stone fruit trees</term><term>Subsequent rflp analysis</term><term>Sugar beet roots</term><term>Variable region</term><term>Virological</term><term>Virological methods</term><term>Woody plants</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: Molecular techniques based on the polymerase chain reaction (PCR) can provide rapid and sensitive diagnosis of plum pox virus (PPV), the causal agent of the devastating `sharka' disease of stone fruit trees. The present study compared routine polymerase chain reaction (PCR) procedures against a new system, PCR-ELISA (Boehringer Mannheim), which enables immunoenzymatic detection of PCR products. The results show that this hybridisation system ensures fast and more sensitive detection of PPV associated with stone fruit trees and herbaceous hosts. Strain-specific capture probes were also designed to identify the two major PPV isolates, D and M, without subsequent restriction fragment length polymorphism analysis of the PCR products. Optimisation of all parameters involved in the PCR-ELISA procedure are discussed and its advantages reported.</div></front></TEI><affiliations><list><country><li>Italie</li></country></list><tree><country name="Italie"><noRegion><name sortKey="Poggi Pollini, C" sort="Poggi Pollini, C" uniqKey="Poggi Pollini C" first="C" last="Poggi Pollini">C. Poggi Pollini</name></noRegion><name sortKey="Bissani, R" sort="Bissani, R" uniqKey="Bissani R" first="R" last="Bissani">R. Bissani</name><name sortKey="Giunchedi, L" sort="Giunchedi, L" uniqKey="Giunchedi L" first="L" last="Giunchedi">L. Giunchedi</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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