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Development of a loop-mediated isothermal amplification method for rapid detection of pigeon circovirus

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Abstract

There are no effective antiviral treatments for pigeon circovirus (PiCV); thus, rapid diagnosis is critical for effective control of the disease caused by this virus. The recent development of a novel LAMP technique that amplifies nucleic acids rapidly with high specificity and sensitivity under isothermal conditions has overcome some of the deficiencies of nucleic-acid-based diagnostic tests. We established a LAMP method for rapid detection of PiCV using two pairs of primers that were designed from PiCV and compared its sensitivity and specificity with that of PCR. Amplification by LAMP was optimal at 63 °C for 60 min. The detection limit was nearly 0.5 pg of PiCV DNA, making it ten times more sensitive than PCR. There was no cross-reaction with porcine circovirus type 2 (PCV2), pigeon Trichomonas gallinae, or pigeon herpesvirus (PHV) under the same conditions. The assay also successfully detected the pathogen DNA in the tissues of infected pigeons. This is the first report indicating that LAMP is a valuable, rapid method of detecting PiCV with high sensitivity and specificity.

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Abbreviations

BIP:

Backward inner primer

FIP:

Forward inner primer

LAMP:

Loop-mediated isothermal amplification

PCR:

Polymerase chain reaction

PiCV:

Pigeon circovirus

PCV2:

Porcine circovirus type 2

PHV:

Pigeon herpesvirus

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Acknowledgements

These studies were supported by Grants to Kuo Pin Chuang from the National Science Council (NSC) and Council of Agriculture of Taiwan.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Veterinary Medicine, National Pingtung University of Science and Technology, Pingtung, 912, Taiwan

    Shinn Shyong Tsai, Yen Li Huang & Tsung Chou Chang

  2. Graduate Institute of Animal Vaccine Technology, National Pingtung University of Science and Technology, Pingtung, 912, Taiwan

    Yeng Ling Chang, Guan Ming Ke, Li Ting Cheng & Kuo Pin Chuang

  3. Institute of Molecular Biology, National Chung Hsing University, Taichung, 250, Taiwan

    Hung Jen Liu

  4. Bureau of Animal and Plant Health Inspection and Quarantine, Council of Agriculture, Executive Yuan, Taipei, 100, Taiwan

    Chwei Jang Chiou & Yao Ching Hsieh

Authors
  1. Shinn Shyong Tsai
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  2. Yeng Ling Chang
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  3. Yen Li Huang
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  4. Hung Jen Liu
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  5. Guan Ming Ke
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  6. Chwei Jang Chiou
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  7. Yao Ching Hsieh
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  8. Tsung Chou Chang
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  9. Li Ting Cheng
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  10. Kuo Pin Chuang
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Correspondence to Kuo Pin Chuang.

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Tsai, S.S., Chang, Y.L., Huang, Y.L. et al. Development of a loop-mediated isothermal amplification method for rapid detection of pigeon circovirus. Arch Virol 159, 921–926 (2014). https://doi.org/10.1007/s00705-013-1906-1

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  • DOI: https://doi.org/10.1007/s00705-013-1906-1

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