Abstract
A novel fluorescent probe-based real-time reverse transcription recombinase polymerase amplification (real-time RT-RPA) assay was developed for rapid detection of highly pathogenic type 2 porcine reproductive and respiratory syndrome virus (HP-PRRSV). The sensitivity analysis showed that the detection limit of RPA was 70 copies of HP-PRRSV RNA/reaction. The real-time RT-RPA highly specific amplified HP-PRRSV with no cross-reaction with classic PRRSV, classic swine fever virus, pseudorabies virus, and foot-and-mouth disease virus. Assessment with 125 clinical samples showed that the developed real-time RT-RPA assay was well correlated with real-time RT-qPCR assays for detection of HP-PRRSV. These results suggest that the developed real-time RT-RPA assay is suitable for rapid detection of HP-PRRSV.
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References
D. Cavanagh, Arch. Virol. 142, 629–633 (1997)
E. Albina, Vet. Microbiol. 55, 309–316 (1997)
G. Wensvoort, C. Terpstra, J.M. Pol, E.A. ter Laak, M. Bloemraad, E.P. de Kluyver, C. Kragten, L. van Buiten, A. den Besten, F. Wagenaar, Vet Q. 13, 121–130 (1991)
K. Tian, X. Yu, T. Zhao, Y. Feng, Z. Cao, C. Wang, Y. Hu, X. Chen, D. Hu, X. Tian, D. Liu, S. Zhang, X. Deng, Y. Ding, L. Yang, Y. Zhang, H. Xiao, M. Qiao, B. Wang, L. Hou, X. Wang, X. Yang, L. Kang, M. Sun, P. Jin, S. Wang, Y. Kitamura, J. Yan, G.F. Gao, PLoS One 2, e526 (2007)
Z. Chai, W. Ma, F. Fu, Y. Lang, W. Wang, G. Tong, Q. Liu, X. Cai, X. Li, Arch. Virol. 158, 407–415 (2013)
K. Yang, Y. Li, Z. Duan, R. Guo, Z. Liu, D. Zhou, F. Yuan, Y. Tian, Gene 531, 199–204 (2013)
X.L. Xiao, H. Wu, Y.G. Yu, B.Z. Cheng, X.Q. Yang, G. Chen, D.M. Liu, X.F. Li, J. Virol. Methods 149, 49–55 (2008)
N.H. Chen, X.Z. Chen, D.M. Hu, X.L. Yu, L.L. Wang, W. Han, J.J. Wu, Z. Cao, C.B. Wang, Q. Zhang, B.Y. Wang, K.G. Tian, J. Virol. Methods 161, 192–198 (2009)
C. Escadafal, O. Faye, A.A. Sall, O. Faye, M. Weidmann, O. Strohmeier, F. von Stetten, J. Drexler, M. Eberhard, M. Niedrig, P. Patel, PLoS Negl Trop Dis. 8, e2730 (2014)
X. Xia, Y. Yu, M. Weidmann, Y. Pan, S. Yan, Y. Wang, PLoS One 9, e104667 (2014)
A. Liljander, M. Yu, E. O’Brien, M. Heller, J.F. Nepper, D.B. Weibel, I. Gluecks, M. Younan, J. Frey, L. Falquet, J. Jores, J. Clin. Microbiol. 53, 2810–2815 (2015)
A. Abd El Wahed, P. Patel, O. Faye, S. Thaloengsok, D. Heidenreich, P. Matangkasombut, K. Manopwisedjaroen, A. Sakuntabhai, A.A. Sall, F.T. Hufert, M. Weidmann, PLoS One 10, e0129682 (2015)
Y. Yang, X. Qin, G. Wang, Y. Zhang, Y. Shang, Z. Zhang, Virol. J. 12, 206 (2015)
M. Euler, Y. Wang, O. Nentwich, O. Piepenburg, F.T. Hufert, M. Weidmann, J. Clin. Virol. 54, 308–312 (2012)
M. Euler, Y. Wang, P. Otto, H. Tomaso, R. Escudero, P. Anda, F.T. Hufert, M. Weidmann, J. Clin. Microbiol. 50, 2234–2238 (2012)
Authors’ contributions
YY, XDQ and ZZ conceived and designed the project. YY performed the study and wrote manuscript. YJS and TC participated in preparation of samples. XDQ and ZZ revised the manuscript. ZZ is the leader of the project. All authors read and approved the final manuscript.
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All the authors declare that they have no competing interests.
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This article does not contain any studies with human participants or animals performed by any of the authors.
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This work was funded by Innovation Fund of the Chinese Academy of Agricultural Sciences (CAAS), China, and the National Natural Science Foundation of China (No. 31572522).
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Yang, Y., Qin, X., Sun, Y. et al. Rapid detection of highly pathogenic porcine reproductive and respiratory syndrome virus by a fluorescent probe-based isothermal recombinase polymerase amplification assay. Virus Genes 52, 883–886 (2016). https://doi.org/10.1007/s11262-016-1378-y
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DOI: https://doi.org/10.1007/s11262-016-1378-y