Bead-based suspension array for simultaneous differential detection of five major swine viruses
Abstract A novel multiplex detection array based on Luminex xMAP technology was developed and validated for simultaneous detection of five major viruses causing swine reproductive diseases. By combining one-step asymmetric multiplex reverse transcription polymerase chain reaction (RT-PCR) with xMAP...
Ausführliche Beschreibung
Autor*in: |
Chen, Ru [verfasserIn] |
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Artikel |
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Sprache: |
Englisch |
Erschienen: |
2015 |
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Anmerkung: |
© Springer-Verlag Berlin Heidelberg 2015 |
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Übergeordnetes Werk: |
Enthalten in: Applied microbiology and biotechnology - Springer Berlin Heidelberg, 1984, 99(2015), 2 vom: Jan., Seite 919-928 |
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Übergeordnetes Werk: |
volume:99 ; year:2015 ; number:2 ; month:01 ; pages:919-928 |
Links: |
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DOI / URN: |
10.1007/s00253-014-6337-8 |
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Katalog-ID: |
OLC2050764537 |
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700 | 1 | |a Song, Chang-Xu |4 aut | |
700 | 1 | |a Li, Yan |4 aut | |
700 | 1 | |a Cao, Yong-Chang |4 aut | |
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10.1007/s00253-014-6337-8 doi (DE-627)OLC2050764537 (DE-He213)s00253-014-6337-8-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Chen, Ru verfasserin aut Bead-based suspension array for simultaneous differential detection of five major swine viruses 2015 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag Berlin Heidelberg 2015 Abstract A novel multiplex detection array based on Luminex xMAP technology was developed and validated for simultaneous detection of five major viruses causing swine reproductive diseases. By combining one-step asymmetric multiplex reverse transcription polymerase chain reaction (RT-PCR) with xMAP bead-based hybridization and flow cytometry analysis, the resulting multiplex assay was capable of detecting single and mixed infections of PRRSV, PCV-2, PRV, CSFV, and PPV in a single reaction. The assay accurately detected and differentiated 23 viral strains used in this study. The low detection limit was determined as 2.2–22 copies/μL (corresponding to 0.5–6.8 fg/μL DNA template) on plasmid constructs containing viral fragments. The intra-assay and inter-assay variances (CV%) were low that ranged from 2.5 to 5.4 % and 4.1 to 7.6 %, respectively. The assay was applied to test field samples and detected single and mixed viral infections. The detection rate was higher than that of uniplex conventional PCR and RT-PCR methods. The detection of PRRSV by the bead-based multiplex assay was comparable with a commercially available real time RT-PCR kit. The test procedure on purified DNA or RNA samples could be completed within 2 h. In conclusion, the bead-based suspension array presented here proved to be a high-throughput practical tool that provided highly specific and sensitive identification of single and multiple infections of five major viruses in pigs and boar semen. xMAP technology Bead-based suspension array Swine virus Multiplex detection Yu, Xiao-Lu aut Gao, Xiao-Bo aut Xue, Cun-Yi aut Song, Chang-Xu aut Li, Yan aut Cao, Yong-Chang aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 99(2015), 2 vom: Jan., Seite 919-928 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:99 year:2015 number:2 month:01 pages:919-928 https://doi.org/10.1007/s00253-014-6337-8 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_70 GBV_ILN_130 GBV_ILN_267 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4082 GBV_ILN_4277 GBV_ILN_4305 AR 99 2015 2 01 919-928 |
spelling |
10.1007/s00253-014-6337-8 doi (DE-627)OLC2050764537 (DE-He213)s00253-014-6337-8-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Chen, Ru verfasserin aut Bead-based suspension array for simultaneous differential detection of five major swine viruses 2015 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag Berlin Heidelberg 2015 Abstract A novel multiplex detection array based on Luminex xMAP technology was developed and validated for simultaneous detection of five major viruses causing swine reproductive diseases. By combining one-step asymmetric multiplex reverse transcription polymerase chain reaction (RT-PCR) with xMAP bead-based hybridization and flow cytometry analysis, the resulting multiplex assay was capable of detecting single and mixed infections of PRRSV, PCV-2, PRV, CSFV, and PPV in a single reaction. The assay accurately detected and differentiated 23 viral strains used in this study. The low detection limit was determined as 2.2–22 copies/μL (corresponding to 0.5–6.8 fg/μL DNA template) on plasmid constructs containing viral fragments. The intra-assay and inter-assay variances (CV%) were low that ranged from 2.5 to 5.4 % and 4.1 to 7.6 %, respectively. The assay was applied to test field samples and detected single and mixed viral infections. The detection rate was higher than that of uniplex conventional PCR and RT-PCR methods. The detection of PRRSV by the bead-based multiplex assay was comparable with a commercially available real time RT-PCR kit. The test procedure on purified DNA or RNA samples could be completed within 2 h. In conclusion, the bead-based suspension array presented here proved to be a high-throughput practical tool that provided highly specific and sensitive identification of single and multiple infections of five major viruses in pigs and boar semen. xMAP technology Bead-based suspension array Swine virus Multiplex detection Yu, Xiao-Lu aut Gao, Xiao-Bo aut Xue, Cun-Yi aut Song, Chang-Xu aut Li, Yan aut Cao, Yong-Chang aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 99(2015), 2 vom: Jan., Seite 919-928 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:99 year:2015 number:2 month:01 pages:919-928 https://doi.org/10.1007/s00253-014-6337-8 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_70 GBV_ILN_130 GBV_ILN_267 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4082 GBV_ILN_4277 GBV_ILN_4305 AR 99 2015 2 01 919-928 |
allfields_unstemmed |
10.1007/s00253-014-6337-8 doi (DE-627)OLC2050764537 (DE-He213)s00253-014-6337-8-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Chen, Ru verfasserin aut Bead-based suspension array for simultaneous differential detection of five major swine viruses 2015 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag Berlin Heidelberg 2015 Abstract A novel multiplex detection array based on Luminex xMAP technology was developed and validated for simultaneous detection of five major viruses causing swine reproductive diseases. By combining one-step asymmetric multiplex reverse transcription polymerase chain reaction (RT-PCR) with xMAP bead-based hybridization and flow cytometry analysis, the resulting multiplex assay was capable of detecting single and mixed infections of PRRSV, PCV-2, PRV, CSFV, and PPV in a single reaction. The assay accurately detected and differentiated 23 viral strains used in this study. The low detection limit was determined as 2.2–22 copies/μL (corresponding to 0.5–6.8 fg/μL DNA template) on plasmid constructs containing viral fragments. The intra-assay and inter-assay variances (CV%) were low that ranged from 2.5 to 5.4 % and 4.1 to 7.6 %, respectively. The assay was applied to test field samples and detected single and mixed viral infections. The detection rate was higher than that of uniplex conventional PCR and RT-PCR methods. The detection of PRRSV by the bead-based multiplex assay was comparable with a commercially available real time RT-PCR kit. The test procedure on purified DNA or RNA samples could be completed within 2 h. In conclusion, the bead-based suspension array presented here proved to be a high-throughput practical tool that provided highly specific and sensitive identification of single and multiple infections of five major viruses in pigs and boar semen. xMAP technology Bead-based suspension array Swine virus Multiplex detection Yu, Xiao-Lu aut Gao, Xiao-Bo aut Xue, Cun-Yi aut Song, Chang-Xu aut Li, Yan aut Cao, Yong-Chang aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 99(2015), 2 vom: Jan., Seite 919-928 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:99 year:2015 number:2 month:01 pages:919-928 https://doi.org/10.1007/s00253-014-6337-8 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_70 GBV_ILN_130 GBV_ILN_267 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4082 GBV_ILN_4277 GBV_ILN_4305 AR 99 2015 2 01 919-928 |
allfieldsGer |
10.1007/s00253-014-6337-8 doi (DE-627)OLC2050764537 (DE-He213)s00253-014-6337-8-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Chen, Ru verfasserin aut Bead-based suspension array for simultaneous differential detection of five major swine viruses 2015 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag Berlin Heidelberg 2015 Abstract A novel multiplex detection array based on Luminex xMAP technology was developed and validated for simultaneous detection of five major viruses causing swine reproductive diseases. By combining one-step asymmetric multiplex reverse transcription polymerase chain reaction (RT-PCR) with xMAP bead-based hybridization and flow cytometry analysis, the resulting multiplex assay was capable of detecting single and mixed infections of PRRSV, PCV-2, PRV, CSFV, and PPV in a single reaction. The assay accurately detected and differentiated 23 viral strains used in this study. The low detection limit was determined as 2.2–22 copies/μL (corresponding to 0.5–6.8 fg/μL DNA template) on plasmid constructs containing viral fragments. The intra-assay and inter-assay variances (CV%) were low that ranged from 2.5 to 5.4 % and 4.1 to 7.6 %, respectively. The assay was applied to test field samples and detected single and mixed viral infections. The detection rate was higher than that of uniplex conventional PCR and RT-PCR methods. The detection of PRRSV by the bead-based multiplex assay was comparable with a commercially available real time RT-PCR kit. The test procedure on purified DNA or RNA samples could be completed within 2 h. In conclusion, the bead-based suspension array presented here proved to be a high-throughput practical tool that provided highly specific and sensitive identification of single and multiple infections of five major viruses in pigs and boar semen. xMAP technology Bead-based suspension array Swine virus Multiplex detection Yu, Xiao-Lu aut Gao, Xiao-Bo aut Xue, Cun-Yi aut Song, Chang-Xu aut Li, Yan aut Cao, Yong-Chang aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 99(2015), 2 vom: Jan., Seite 919-928 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:99 year:2015 number:2 month:01 pages:919-928 https://doi.org/10.1007/s00253-014-6337-8 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_70 GBV_ILN_130 GBV_ILN_267 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4082 GBV_ILN_4277 GBV_ILN_4305 AR 99 2015 2 01 919-928 |
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10.1007/s00253-014-6337-8 doi (DE-627)OLC2050764537 (DE-He213)s00253-014-6337-8-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Chen, Ru verfasserin aut Bead-based suspension array for simultaneous differential detection of five major swine viruses 2015 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag Berlin Heidelberg 2015 Abstract A novel multiplex detection array based on Luminex xMAP technology was developed and validated for simultaneous detection of five major viruses causing swine reproductive diseases. By combining one-step asymmetric multiplex reverse transcription polymerase chain reaction (RT-PCR) with xMAP bead-based hybridization and flow cytometry analysis, the resulting multiplex assay was capable of detecting single and mixed infections of PRRSV, PCV-2, PRV, CSFV, and PPV in a single reaction. The assay accurately detected and differentiated 23 viral strains used in this study. The low detection limit was determined as 2.2–22 copies/μL (corresponding to 0.5–6.8 fg/μL DNA template) on plasmid constructs containing viral fragments. The intra-assay and inter-assay variances (CV%) were low that ranged from 2.5 to 5.4 % and 4.1 to 7.6 %, respectively. The assay was applied to test field samples and detected single and mixed viral infections. The detection rate was higher than that of uniplex conventional PCR and RT-PCR methods. The detection of PRRSV by the bead-based multiplex assay was comparable with a commercially available real time RT-PCR kit. The test procedure on purified DNA or RNA samples could be completed within 2 h. In conclusion, the bead-based suspension array presented here proved to be a high-throughput practical tool that provided highly specific and sensitive identification of single and multiple infections of five major viruses in pigs and boar semen. xMAP technology Bead-based suspension array Swine virus Multiplex detection Yu, Xiao-Lu aut Gao, Xiao-Bo aut Xue, Cun-Yi aut Song, Chang-Xu aut Li, Yan aut Cao, Yong-Chang aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 99(2015), 2 vom: Jan., Seite 919-928 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:99 year:2015 number:2 month:01 pages:919-928 https://doi.org/10.1007/s00253-014-6337-8 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_70 GBV_ILN_130 GBV_ILN_267 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4082 GBV_ILN_4277 GBV_ILN_4305 AR 99 2015 2 01 919-928 |
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Enthalten in Applied microbiology and biotechnology 99(2015), 2 vom: Jan., Seite 919-928 volume:99 year:2015 number:2 month:01 pages:919-928 |
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Chen, Ru @@aut@@ Yu, Xiao-Lu @@aut@@ Gao, Xiao-Bo @@aut@@ Xue, Cun-Yi @@aut@@ Song, Chang-Xu @@aut@@ Li, Yan @@aut@@ Cao, Yong-Chang @@aut@@ |
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Bead-based suspension array for simultaneous differential detection of five major swine viruses |
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Abstract A novel multiplex detection array based on Luminex xMAP technology was developed and validated for simultaneous detection of five major viruses causing swine reproductive diseases. By combining one-step asymmetric multiplex reverse transcription polymerase chain reaction (RT-PCR) with xMAP bead-based hybridization and flow cytometry analysis, the resulting multiplex assay was capable of detecting single and mixed infections of PRRSV, PCV-2, PRV, CSFV, and PPV in a single reaction. The assay accurately detected and differentiated 23 viral strains used in this study. The low detection limit was determined as 2.2–22 copies/μL (corresponding to 0.5–6.8 fg/μL DNA template) on plasmid constructs containing viral fragments. The intra-assay and inter-assay variances (CV%) were low that ranged from 2.5 to 5.4 % and 4.1 to 7.6 %, respectively. The assay was applied to test field samples and detected single and mixed viral infections. The detection rate was higher than that of uniplex conventional PCR and RT-PCR methods. The detection of PRRSV by the bead-based multiplex assay was comparable with a commercially available real time RT-PCR kit. The test procedure on purified DNA or RNA samples could be completed within 2 h. In conclusion, the bead-based suspension array presented here proved to be a high-throughput practical tool that provided highly specific and sensitive identification of single and multiple infections of five major viruses in pigs and boar semen. © Springer-Verlag Berlin Heidelberg 2015 |
abstractGer |
Abstract A novel multiplex detection array based on Luminex xMAP technology was developed and validated for simultaneous detection of five major viruses causing swine reproductive diseases. By combining one-step asymmetric multiplex reverse transcription polymerase chain reaction (RT-PCR) with xMAP bead-based hybridization and flow cytometry analysis, the resulting multiplex assay was capable of detecting single and mixed infections of PRRSV, PCV-2, PRV, CSFV, and PPV in a single reaction. The assay accurately detected and differentiated 23 viral strains used in this study. The low detection limit was determined as 2.2–22 copies/μL (corresponding to 0.5–6.8 fg/μL DNA template) on plasmid constructs containing viral fragments. The intra-assay and inter-assay variances (CV%) were low that ranged from 2.5 to 5.4 % and 4.1 to 7.6 %, respectively. The assay was applied to test field samples and detected single and mixed viral infections. The detection rate was higher than that of uniplex conventional PCR and RT-PCR methods. The detection of PRRSV by the bead-based multiplex assay was comparable with a commercially available real time RT-PCR kit. The test procedure on purified DNA or RNA samples could be completed within 2 h. In conclusion, the bead-based suspension array presented here proved to be a high-throughput practical tool that provided highly specific and sensitive identification of single and multiple infections of five major viruses in pigs and boar semen. © Springer-Verlag Berlin Heidelberg 2015 |
abstract_unstemmed |
Abstract A novel multiplex detection array based on Luminex xMAP technology was developed and validated for simultaneous detection of five major viruses causing swine reproductive diseases. By combining one-step asymmetric multiplex reverse transcription polymerase chain reaction (RT-PCR) with xMAP bead-based hybridization and flow cytometry analysis, the resulting multiplex assay was capable of detecting single and mixed infections of PRRSV, PCV-2, PRV, CSFV, and PPV in a single reaction. The assay accurately detected and differentiated 23 viral strains used in this study. The low detection limit was determined as 2.2–22 copies/μL (corresponding to 0.5–6.8 fg/μL DNA template) on plasmid constructs containing viral fragments. The intra-assay and inter-assay variances (CV%) were low that ranged from 2.5 to 5.4 % and 4.1 to 7.6 %, respectively. The assay was applied to test field samples and detected single and mixed viral infections. The detection rate was higher than that of uniplex conventional PCR and RT-PCR methods. The detection of PRRSV by the bead-based multiplex assay was comparable with a commercially available real time RT-PCR kit. The test procedure on purified DNA or RNA samples could be completed within 2 h. In conclusion, the bead-based suspension array presented here proved to be a high-throughput practical tool that provided highly specific and sensitive identification of single and multiple infections of five major viruses in pigs and boar semen. © Springer-Verlag Berlin Heidelberg 2015 |
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