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Development of RT-LAMP and real-time RT-PCR assays for the rapid detection of the new duck Tembusu-like BYD virus

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Abstract

A new duck Tembusu virus (TMUV), also known as BYD virus, has been identified as the causative agent for the emerging duck egg-drop syndrome in mainland China. The rapid spread and wide distribution of the new TMUV in mainland China result in heavy loss to the poultry industry and pose great threats to public health. Rapid and sensitive detection methods are critical for prevention and control of TMUV infections. In this study, a reverse-transcription loop-mediated isothermal amplification assay (RT-LAMP) and an SYBR Green-I–based real-time RT-PCR assay specific for the duck TMUV were developed and validated with laboratory and field samples, respectively. The detection limits were 1 × 10−4 and 1 × 10−3 PFU per reaction for the RT-LAMP assay and real-time RT-PCR assay, respectively. The specificities were analyzed with other related members of the genus Flavivirus, and no cross-reaction was observed. Furthermore, both assays were evaluated with field samples, and they exhibited high sensitivity and specificity. In addition, the real-time RT-PCR assay worked well in viral load analysis, which revealed that the spleen may be the primary target for the replication of new duck TMUV in ducks. The TMUV-specific RT-LAMP and real-time RT-PCR assays will provide useful tools for the diagnosis and epidemiological surveillance of TMUV infection.

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Acknowledgments

This work was supported, in part, by the Major Special Program of National Science and Technology of China (No.2009ZX10004-204), and the National Natural Science Foundation of China (No. 30972613, No. 81000722). CFQ was supported by Beijing Nova Program of Science and Technology (No.2010B041).

Conflict of interests

The authors have declared that no competing interests exist.

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Authors and Affiliations

  1. State Key Laboratory of Pathogen and Biosecurity, Department of Virology, Beijing Institute of Microbiology and Epidemiology, No. 20 Dongda Street, Fengtai District, 100071, Beijing, China

    Tao Jiang, Juan Liu, Yong-Qiang Deng, Li-Juan Xu, Zhi-Hui Liu, Xiao-Feng Li, Xue-Dong Yu, Shun-Ya Zhu, E-De Qin & Cheng-Feng Qin

  2. Department of Transfusion Medicine, PLA Air Force General Hospital, 100142, Beijing, China

    Juan Liu

  3. Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, 100193, Beijing, China

    Jing-Liang Su

  4. College of Veterinary Medicine, Xinjiang Agricultural University, 830052, Urumqi, China

    Zhi-Hui Liu

  5. CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, 100101, Beijing, China

    George Fu Gao

Authors
  1. Tao Jiang
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  2. Juan Liu
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  3. Yong-Qiang Deng
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  4. Jing-Liang Su
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  5. Li-Juan Xu
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  6. Zhi-Hui Liu
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  7. Xiao-Feng Li
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  8. Xue-Dong Yu
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  9. Shun-Ya Zhu
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  10. George Fu Gao
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  11. E-De Qin
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  12. Cheng-Feng Qin
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Corresponding author

Correspondence to Cheng-Feng Qin.

Additional information

T. Jiang and J. Liu contributed equally to this work.

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Jiang, T., Liu, J., Deng, YQ. et al. Development of RT-LAMP and real-time RT-PCR assays for the rapid detection of the new duck Tembusu-like BYD virus. Arch Virol 157, 2273–2280 (2012). https://doi.org/10.1007/s00705-012-1431-7

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  • DOI: https://doi.org/10.1007/s00705-012-1431-7

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