A one‐step molecular biology method for simple and rapid detection of grass carp Ctenopharyngodon idella reovirus (GCRV) HZ08 strain
Identifieur interne : 000303 ( Istex/Checkpoint ); précédent : 000302; suivant : 000304A one‐step molecular biology method for simple and rapid detection of grass carp Ctenopharyngodon idella reovirus (GCRV) HZ08 strain
Auteurs : W. W. Zeng [République populaire de Chine] ; Q. Wang [République populaire de Chine] ; Y. Y. Wang [République populaire de Chine] ; D. H. Xu [États-Unis] ; S. Q. Wu [République populaire de Chine]Source :
- Journal of Fish Biology [ 0022-1112 ] ; 2013-05.
English descriptors
- Teeft :
- Acta hydrobiologic sinica, Assay, Atcc number, Authors journal, British isles, Cell line, Cellular probes, Chinese academy, Clinical samples, Ctenopharyngodon, Ctenopharyngodon idella, Ctenopharyngodon idella reovirus, Cyprinus carpio carpio, Detection limit, Disease virus, Diseased, Early detection, Early stages, Eiken chemical, Elution buffer, Fish biology, Fisheries society, Fishery, Fishery sciences, Gcrv, Gcrv strain, Gsrv, Haemorrhagic disease, Hemorrhagic virus, Higher sensitivity, Idella, Ihnv, Isknv, Isothermal, Isothermal journal, Jiangxi province, Loop primers, Negative control, Notomi, Nucleotide sequences, Optimal temperature, Optimum reaction temperature, Original publication, Polymerase, Polymerase chain reaction, Positive control, Positive samples, Primer, Rapid detection, Reaction volume, Reovirus, Science fund, Sensitive detection, Simple assay, Subtype infection, Virological, Virological methods, Virus isolation, Wang, Ytav, Zeng, Zhang.
Abstract
Six reverse‐transcription loop‐mediated isothermal amplification (RT‐LAMP) primers designed against conserved regions of segment 6 (s6) gene were used for the detection of grass carp Ctenopharyngodon idella reovirus (GCRV) HZ08 subtype. The entire amplification could be completed within 40 min at 62·3° C. The RT‐LAMP showed higher sensitivity than reverse‐transcription polymerase chain reaction (RT‐PCR). The RNA detection limit was 10 copies µl−1 for RT‐LAMP assay and 100 copies µl−1 for conventional RT‐PCR. In specificity tests, no cross‐reactivity was detected in other viruses from common aquatic animals. In addition, the reaction results can be visualized by using calcein fluorescent dye. Furthermore, a total of 86 samples were tested by RT‐LAMP, RT‐PCR and virus isolation. The results demonstrated that all 54 specimens identified as positive by virus isolation were also positive when detected by RT‐LAMP. Seven out of 54 samples, however, were misidentified by RT‐PCR. The RT‐LAMP method is more accurate than conventional RT‐PCR. The results indicate that RT‐LAMP has potential as a simple and rapid diagnosis technique for the detection of GCRV HZ08 subtype infection.
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DOI: 10.1111/jfb.12088
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Zeng</name><affiliation wicri:level="3"><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences, 510380, Guangzhou</wicri:regionArea><placeName><settlement type="city">Jiangmen</settlement><region type="province">Guangdong</region></placeName></affiliation></author><author><name sortKey="Wang, Q" sort="Wang, Q" uniqKey="Wang Q" first="Q." last="Wang">Q. Wang</name><affiliation wicri:level="3"><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences, 510380, Guangzhou</wicri:regionArea><placeName><settlement type="city">Jiangmen</settlement><region type="province">Guangdong</region></placeName></affiliation></author><author><name sortKey="Wang, Y Y" sort="Wang, Y Y" uniqKey="Wang Y" first="Y. Y." last="Wang">Y. Y. 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Wu</name><affiliation wicri:level="3"><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences, 510380, Guangzhou</wicri:regionArea><placeName><settlement type="city">Jiangmen</settlement><region type="province">Guangdong</region></placeName></affiliation><affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j" type="main">Journal of Fish Biology</title><title level="j" type="alt">JOURNAL OF FISH BIOLOGY</title><idno type="ISSN">0022-1112</idno><idno type="eISSN">1095-8649</idno><imprint><biblScope unit="vol">82</biblScope><biblScope unit="issue">5</biblScope><biblScope unit="page" from="1545">1545</biblScope><biblScope unit="page" to="1555">1555</biblScope><biblScope unit="page-count">11</biblScope><publisher>Blackwell Publishing Ltd</publisher><pubPlace>Oxford, UK</pubPlace><date type="published" when="2013-05">2013-05</date></imprint><idno type="ISSN">0022-1112</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0022-1112</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Acta hydrobiologic sinica</term><term>Assay</term><term>Atcc number</term><term>Authors journal</term><term>British isles</term><term>Cell line</term><term>Cellular probes</term><term>Chinese academy</term><term>Clinical samples</term><term>Ctenopharyngodon</term><term>Ctenopharyngodon idella</term><term>Ctenopharyngodon idella reovirus</term><term>Cyprinus carpio carpio</term><term>Detection limit</term><term>Disease virus</term><term>Diseased</term><term>Early detection</term><term>Early stages</term><term>Eiken chemical</term><term>Elution buffer</term><term>Fish biology</term><term>Fisheries society</term><term>Fishery</term><term>Fishery sciences</term><term>Gcrv</term><term>Gcrv strain</term><term>Gsrv</term><term>Haemorrhagic disease</term><term>Hemorrhagic virus</term><term>Higher sensitivity</term><term>Idella</term><term>Ihnv</term><term>Isknv</term><term>Isothermal</term><term>Isothermal journal</term><term>Jiangxi province</term><term>Loop primers</term><term>Negative control</term><term>Notomi</term><term>Nucleotide sequences</term><term>Optimal temperature</term><term>Optimum reaction temperature</term><term>Original publication</term><term>Polymerase</term><term>Polymerase chain reaction</term><term>Positive control</term><term>Positive samples</term><term>Primer</term><term>Rapid detection</term><term>Reaction volume</term><term>Reovirus</term><term>Science fund</term><term>Sensitive detection</term><term>Simple assay</term><term>Subtype infection</term><term>Virological</term><term>Virological methods</term><term>Virus isolation</term><term>Wang</term><term>Ytav</term><term>Zeng</term><term>Zhang</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract">Six reverse‐transcription loop‐mediated isothermal amplification (RT‐LAMP) primers designed against conserved regions of segment 6 (s6) gene were used for the detection of grass carp Ctenopharyngodon idella reovirus (GCRV) HZ08 subtype. The entire amplification could be completed within 40 min at 62·3° C. The RT‐LAMP showed higher sensitivity than reverse‐transcription polymerase chain reaction (RT‐PCR). The RNA detection limit was 10 copies µl−1 for RT‐LAMP assay and 100 copies µl−1 for conventional RT‐PCR. In specificity tests, no cross‐reactivity was detected in other viruses from common aquatic animals. In addition, the reaction results can be visualized by using calcein fluorescent dye. Furthermore, a total of 86 samples were tested by RT‐LAMP, RT‐PCR and virus isolation. The results demonstrated that all 54 specimens identified as positive by virus isolation were also positive when detected by RT‐LAMP. Seven out of 54 samples, however, were misidentified by RT‐PCR. The RT‐LAMP method is more accurate than conventional RT‐PCR. The results indicate that RT‐LAMP has potential as a simple and rapid diagnosis technique for the detection of GCRV HZ08 subtype infection.</div></front></TEI><affiliations><list><country><li>République populaire de Chine</li><li>États-Unis</li></country><region><li>Guangdong</li></region><settlement><li>Jiangmen</li></settlement></list><tree><country name="République populaire de Chine"><region name="Guangdong"><name sortKey="Zeng, W W" sort="Zeng, W W" uniqKey="Zeng W" first="W. W." last="Zeng">W. W. Zeng</name></region><name sortKey="Wang, Q" sort="Wang, Q" uniqKey="Wang Q" first="Q." last="Wang">Q. Wang</name><name sortKey="Wang, Y Y" sort="Wang, Y Y" uniqKey="Wang Y" first="Y. Y." last="Wang">Y. Y. Wang</name><name sortKey="Wu, S Q" sort="Wu, S Q" uniqKey="Wu S" first="S. Q." last="Wu">S. Q. Wu</name></country><country name="États-Unis"><noRegion><name sortKey="Xu, D H" sort="Xu, D H" uniqKey="Xu D" first="D. H." last="Xu">D. H. Xu</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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