Rapid detection of the common avian leukosis virus subgroups by real-time loop-mediated isothermal amplification
Identifieur interne : 000922 ( Main/Exploration ); précédent : 000921; suivant : 000923Rapid detection of the common avian leukosis virus subgroups by real-time loop-mediated isothermal amplification
Auteurs : Hao Peng [République populaire de Chine] ; Lili Qin [République populaire de Chine] ; Yuyu Bi [République populaire de Chine] ; Peikun Wang [République populaire de Chine] ; Guangzhen Zou [République populaire de Chine] ; Jun Li [République populaire de Chine] ; Yongli Yang [République populaire de Chine] ; Xingfu Zhong [République populaire de Chine] ; Ping Wei [République populaire de Chine]Source :
- Virology Journal [ 1743-422X ] ; 2015.
Abstract
Subgroups A, B, E and J are the major subgroups of avian leukosis virus (ALV) infecting chickens. ALV infection has become endemic in China and has a significant negative effect on the poultry industry. Consequently, there is an urgent need for a specific, sensitive and rapid method for diagnosis and eradication of ALV. Therefore, we developed a simple and rapid real-time loop-mediated isothermal amplification (LAMP) reaction for the timely detection of the common ALV subgroups, whereby the amplification can be obtained in 35 min under isothermal conditions at 63 °C, ability to specific, sensitive and rapid detect all the common ALV subgroups.
A set of four specific primers was designed to target the sequences of the
The results from specificity of the LAMP assay showed that only target ALVs DNA was amplified. The LAMP assay demonstrated a sensitivity of 20 copies/reaction of ALV DNA, which was 10 times higher than the conventional PCR measurement. To further evaluate the reliability of the method, the assay was evaluated with ALV DNA from a panel of 81 clinical samples suspected of ALV infection. The results verify that the LAMP method was more sensitive than the conventional PCR and virus isolation method.
In conclusion, the developed LAMP assay was a simple, inexpensive, sensitive method for the rapid detection of the most common subgroups of ALV, and it provided a useful and practical tool in the eradication program for ALV in the poultry industry.
Url:
DOI: 10.1186/s12985-015-0430-1
PubMed: 26596553
PubMed Central: 4657318
Affiliations:
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Chine</country><wicri:regionArea>Institute for Poultry Science and Health, Guangxi University, Nanning</wicri:regionArea><wicri:noRegion>Nanning</wicri:noRegion></affiliation></author><author><name sortKey="Zhong, Xingfu" sort="Zhong, Xingfu" uniqKey="Zhong X" first="Xingfu" last="Zhong">Xingfu Zhong</name><affiliation wicri:level="1"><nlm:aff id="Aff1">Institute for Poultry Science and Health, Guangxi University, Nanning, 530004 China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Institute for Poultry Science and Health, Guangxi University, Nanning</wicri:regionArea><wicri:noRegion>Nanning</wicri:noRegion></affiliation></author><author><name sortKey="Wei, Ping" sort="Wei, Ping" uniqKey="Wei P" first="Ping" last="Wei">Ping Wei</name><affiliation wicri:level="1"><nlm:aff id="Aff1">Institute for Poultry Science and Health, Guangxi University, Nanning, 530004 China</nlm:aff><country xml:lang="fr">République populaire de 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Laboratory of Animal Vaccines and Diagnostics, Guangxi Veterinary Research Institute, 51 Youai Bei Road, Nanning, 530001 China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Guangxi Key Laboratory of Animal Vaccines and Diagnostics, Guangxi Veterinary Research Institute, 51 Youai Bei Road, Nanning</wicri:regionArea><wicri:noRegion>Nanning</wicri:noRegion></affiliation></author><author><name sortKey="Qin, Lili" sort="Qin, Lili" uniqKey="Qin L" first="Lili" last="Qin">Lili Qin</name><affiliation wicri:level="1"><nlm:aff id="Aff1">Institute for Poultry Science and Health, Guangxi University, Nanning, 530004 China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Institute for Poultry Science and Health, Guangxi University, Nanning</wicri:regionArea><wicri:noRegion>Nanning</wicri:noRegion></affiliation></author><author><name sortKey="Bi, Yuyu" sort="Bi, Yuyu" uniqKey="Bi Y" first="Yuyu" last="Bi">Yuyu Bi</name><affiliation wicri:level="1"><nlm:aff id="Aff1">Institute for Poultry Science and Health, Guangxi University, Nanning, 530004 China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Institute for Poultry Science and Health, Guangxi University, Nanning</wicri:regionArea><wicri:noRegion>Nanning</wicri:noRegion></affiliation></author><author><name sortKey="Wang, Peikun" sort="Wang, Peikun" uniqKey="Wang P" first="Peikun" last="Wang">Peikun Wang</name><affiliation wicri:level="1"><nlm:aff id="Aff1">Institute for Poultry Science and Health, Guangxi University, Nanning, 530004 China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Institute for Poultry Science and Health, Guangxi University, Nanning</wicri:regionArea><wicri:noRegion>Nanning</wicri:noRegion></affiliation></author><author><name sortKey="Zou, Guangzhen" sort="Zou, Guangzhen" uniqKey="Zou G" first="Guangzhen" last="Zou">Guangzhen Zou</name><affiliation wicri:level="1"><nlm:aff id="Aff1">Institute for Poultry Science and Health, Guangxi University, Nanning, 530004 China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Institute for Poultry Science and Health, Guangxi University, Nanning</wicri:regionArea><wicri:noRegion>Nanning</wicri:noRegion></affiliation></author><author><name sortKey="Li, Jun" sort="Li, Jun" uniqKey="Li J" first="Jun" last="Li">Jun Li</name><affiliation wicri:level="1"><nlm:aff id="Aff2">Guangxi Key Laboratory of Animal Vaccines and Diagnostics, Guangxi Veterinary Research Institute, 51 Youai Bei Road, Nanning, 530001 China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Guangxi Key Laboratory of Animal Vaccines and Diagnostics, Guangxi Veterinary Research Institute, 51 Youai Bei Road, Nanning</wicri:regionArea><wicri:noRegion>Nanning</wicri:noRegion></affiliation></author><author><name sortKey="Yang, Yongli" sort="Yang, Yongli" uniqKey="Yang Y" first="Yongli" last="Yang">Yongli Yang</name><affiliation wicri:level="1"><nlm:aff id="Aff1">Institute for Poultry Science and Health, Guangxi University, Nanning, 530004 China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Institute for Poultry Science and Health, Guangxi University, Nanning</wicri:regionArea><wicri:noRegion>Nanning</wicri:noRegion></affiliation></author><author><name sortKey="Zhong, Xingfu" sort="Zhong, Xingfu" uniqKey="Zhong X" first="Xingfu" last="Zhong">Xingfu Zhong</name><affiliation wicri:level="1"><nlm:aff id="Aff1">Institute for Poultry Science and Health, Guangxi University, Nanning, 530004 China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Institute for Poultry Science and Health, Guangxi University, Nanning</wicri:regionArea><wicri:noRegion>Nanning</wicri:noRegion></affiliation></author><author><name sortKey="Wei, Ping" sort="Wei, Ping" uniqKey="Wei P" first="Ping" last="Wei">Ping Wei</name><affiliation wicri:level="1"><nlm:aff id="Aff1">Institute for Poultry Science and Health, Guangxi University, Nanning, 530004 China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Institute for Poultry Science and Health, Guangxi University, Nanning</wicri:regionArea><wicri:noRegion>Nanning</wicri:noRegion></affiliation></author></analytic><series><title level="j">Virology Journal</title><idno type="eISSN">1743-422X</idno><imprint><date when="2015">2015</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><sec><title>Background</title><p>Subgroups A, B, E and J are the major subgroups of avian leukosis virus (ALV) infecting chickens. ALV infection has become endemic in China and has a significant negative effect on the poultry industry. Consequently, there is an urgent need for a specific, sensitive and rapid method for diagnosis and eradication of ALV. Therefore, we developed a simple and rapid real-time loop-mediated isothermal amplification (LAMP) reaction for the timely detection of the common ALV subgroups, whereby the amplification can be obtained in 35 min under isothermal conditions at 63 °C, ability to specific, sensitive and rapid detect all the common ALV subgroups.</p></sec><sec><title>Methods</title><p>A set of four specific primers was designed to target the sequences of the <italic>pol</italic> gene of ALV, and the loop-mediated isothermal amplification (LAMP) assay were developed and compared with PCR and virus isolation methods.</p></sec><sec><title>Results</title><p>The results from specificity of the LAMP assay showed that only target ALVs DNA was amplified. The LAMP assay demonstrated a sensitivity of 20 copies/reaction of ALV DNA, which was 10 times higher than the conventional PCR measurement. To further evaluate the reliability of the method, the assay was evaluated with ALV DNA from a panel of 81 clinical samples suspected of ALV infection. The results verify that the LAMP method was more sensitive than the conventional PCR and virus isolation method.</p></sec><sec><title>Conclusion</title><p>In conclusion, the developed LAMP assay was a simple, inexpensive, sensitive method for the rapid detection of the most common subgroups of ALV, and it provided a useful and practical tool in the eradication program for ALV in the poultry industry.</p></sec></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Li, X" uniqKey="Li X">X Li</name></author><author><name sortKey="Dong, X" uniqKey="Dong X">X Dong</name></author><author><name sortKey="Sun, X" uniqKey="Sun X">X Sun</name></author><author><name sortKey="Li, W" uniqKey="Li W">W Li</name></author><author><name sortKey="Zhao, P" uniqKey="Zhao P">P Zhao</name></author><author><name sortKey="Cui, Z" uniqKey="Cui Z">Z Cui</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Salter, Dw" uniqKey="Salter 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Liu</name></author></analytic></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Cui, Z" uniqKey="Cui Z">Z Cui</name></author><author><name sortKey="Du, Y" uniqKey="Du Y">Y Du</name></author><author><name sortKey="Zhang, Z" uniqKey="Zhang Z">Z Zhang</name></author><author><name sortKey="Silva, Rf" uniqKey="Silva R">RF Silva</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Smith, Lm" uniqKey="Smith L">LM Smith</name></author><author><name sortKey="Brown, Sr" uniqKey="Brown S">SR Brown</name></author><author><name sortKey="Howes, K" uniqKey="Howes K">K Howes</name></author><author><name sortKey="Mcleod, S" uniqKey="Mcleod S">S McLeod</name></author><author><name sortKey="Arshad, Ss" uniqKey="Arshad S">SS Arshad</name></author><author><name sortKey="Barron, Gs" uniqKey="Barron G">GS Barron</name></author></analytic></biblStruct><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Kuboki, N" uniqKey="Kuboki N">N Kuboki</name></author><author><name sortKey="Inoue, N" uniqKey="Inoue N">N Inoue</name></author><author><name sortKey="Sakurai, T" uniqKey="Sakurai T">T Sakurai</name></author><author><name sortKey="Cello, Fd" uniqKey="Cello F">FD Cello</name></author><author><name sortKey="Grab, Dj" uniqKey="Grab D">DJ Grab</name></author><author><name sortKey="Suzuki, H" uniqKey="Suzuki H">H Suzuki</name></author></analytic></biblStruct><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Zavala, G" uniqKey="Zavala G">G Zavala</name></author><author><name sortKey="Cheng, S" uniqKey="Cheng S">S Cheng</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list><country><li>République populaire de Chine</li></country></list><tree><country name="République populaire de Chine"><noRegion><name sortKey="Peng, Hao" sort="Peng, Hao" uniqKey="Peng H" first="Hao" last="Peng">Hao Peng</name></noRegion><name sortKey="Bi, Yuyu" sort="Bi, Yuyu" uniqKey="Bi Y" first="Yuyu" last="Bi">Yuyu Bi</name><name sortKey="Li, Jun" sort="Li, Jun" uniqKey="Li J" first="Jun" last="Li">Jun Li</name><name sortKey="Peng, Hao" sort="Peng, Hao" uniqKey="Peng H" first="Hao" last="Peng">Hao Peng</name><name sortKey="Qin, Lili" sort="Qin, Lili" uniqKey="Qin L" first="Lili" last="Qin">Lili Qin</name><name sortKey="Wang, Peikun" sort="Wang, Peikun" uniqKey="Wang P" first="Peikun" last="Wang">Peikun Wang</name><name sortKey="Wei, Ping" sort="Wei, Ping" uniqKey="Wei P" first="Ping" last="Wei">Ping Wei</name><name sortKey="Yang, Yongli" sort="Yang, Yongli" uniqKey="Yang Y" first="Yongli" last="Yang">Yongli Yang</name><name sortKey="Zhong, Xingfu" sort="Zhong, Xingfu" uniqKey="Zhong X" first="Xingfu" last="Zhong">Xingfu Zhong</name><name sortKey="Zou, Guangzhen" sort="Zou, Guangzhen" uniqKey="Zou G" first="Guangzhen" last="Zou">Guangzhen Zou</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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