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Development of a loop-mediated isothermal amplification assay for porcine circovirus type 2

Virologica Sinica volume 26, pages 214–220 (2011)Cite this article

Abstract

In this study, the loop-mediated isothermal amplification (LAMP) method was used to develop a rapid and simple detection system for porcine circovirus type 2 (PCV2). According to the PCV2 sequences published in GenBank, multiple LAMP primers were designed targeting conserved sequences of PCV2. Using the DNA extracted from PCV2 isolates HUN-09 and SD-09 as the template, LAMP reactions in a PCV2 LAMP system was performed, the amplification products were detected by adding SYBR Green I and could be observed directly by the naked eye. The results showed highly-efficient and specific amplification in 30 min at 63°C with a LAMP real-time turbidimeter. Furthermore, PCV2 DNA templates, with a detection limit of 5.5×10−5 ng of nucleic acid, indicated that this assay was highly sensitive. The results obtained with the naked eye after SYBR Green I staining were consistent with those detected by the real-time turbidimeter, showing the potential simplicity of interpretation of the assay results. The LAMP assay appeared to have greater accuracy than PCR and virus isolation for the analysis of 18 clinical samples. In addition it offers higher specificity and sensitivity, shorter reaction times and simpler procedures than the currently available methods of PCV2 detection. It is therefore a promising tool for the effective and efficient detection of PCV2.

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Authors and Affiliations

  1. China Institute of Veterinary Drug Control, Beijing, 100081, China

    Ye-bing Liu, Lei Zhang, Qin-hong Xue, Yi-bao Ning & Zhi-gang Zhang

  2. College of Veterinary Medicine, Northwest Agriculture & Forestry University, Yangling, 712100, China

    Lei Zhang

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  1. Ye-bing Liu
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  2. Lei Zhang
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  3. Qin-hong Xue
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  4. Yi-bao Ning
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  5. Zhi-gang Zhang
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Correspondence to Ye-bing Liu.

Additional information

Foundation item: Fifteenth National Science and Technology Support Program of China (2007BAD86B04-4)

These authors contributed equally.

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Liu, Yb., Zhang, L., Xue, Qh. et al. Development of a loop-mediated isothermal amplification assay for porcine circovirus type 2. Virol. Sin. 26, 214–220 (2011). https://doi.org/10.1007/s12250-011-3169-x

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  • DOI: https://doi.org/10.1007/s12250-011-3169-x

Key words

  • Porcine circovirus type 2 (PCV2)
  • Loop-mediated isothermal amplification (LAMP)
  • Virus detection