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An improved reverse transcription loop-mediated isothermal amplification assay for sensitive and specific detection of Newcastle disease virus

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Abstract

Five of 21 different Newcastle disease virus (NDV) strains isolated from China in 2007 gave false-negative results when detected with a previously reported loop-mediated isothermal amplification (LAMP) method, and mismatches were found between the target gene and LAMP primers. Therefore, an improved, sensitive, specific, and accelerated one-step reverse transcription (RT)-LAMP assay with degenerate primers was developed. Our data demonstrated that the improved RT-LAMP assay detected all 21 NDV isolates, had no cross-reaction with three other avian viruses, could be performed in 50 min less time, was 5-fold more sensitive than the previous LAMP assay, and achieved 96.8% sensitivity with 62 samples, including 30 field clinical samples, 24 experimentally infected samples, and 8 experimentally negative samples. Therefore, the improved RT-LAMP assay is a simple, rapid, and cost-effective method that is practical for less well-equipped laboratories and in the field.

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Acknowledgments

This work was supported by the grants from State Public Industry Scientific Research Programs (nyhyzx07-038) and Major Programs of Science Technology Strategic Plan (2007A020400006) of Guangdong, People's Republic of China. We thank Dr. George D. Liu for kindly revising the manuscript.

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Authors and Affiliations

  1. State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-Sen University, 510006, Guangzhou, People’s Republic of China

    Qiang Li, Chunyi Xue & Yongchang Cao

  2. Guangdong Wen’s Food Co. Ltd, 527400, Xinxing, Guangdong, People’s Republic of China

    Jianping Qin & Qingfeng Zhou

  3. College of Animal Science, South China Agricultural University, 510642, Guangzhou, People’s Republic of China

    Feng Chen & Yingzuo Bi

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  1. Qiang Li
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  2. Chunyi Xue
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  3. Jianping Qin
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  4. Qingfeng Zhou
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  5. Feng Chen
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  6. Yingzuo Bi
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  7. Yongchang Cao
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Correspondence to Yongchang Cao.

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Li, Q., Xue, C., Qin, J. et al. An improved reverse transcription loop-mediated isothermal amplification assay for sensitive and specific detection of Newcastle disease virus. Arch Virol 154, 1433–1440 (2009). https://doi.org/10.1007/s00705-009-0464-z

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  • DOI: https://doi.org/10.1007/s00705-009-0464-z

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