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A real-time PCR to detect and analyze virulent EMCV loads in sows and piglets

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Abstract

A real-time polymerase chain reaction with SYBR Green was developed for the detection and quantification of encephalomyocarditis virus (EMCV) in porcine tissues; the method uses two primers specific for the 3D gene. The detection limit of this assay was 22 gene copies/reaction, equivalent to 0.001 TCID50/ml. The assay was linear over a 107 dilution range of template concentrations and was specific for EMCV; it did not amplify other porcine pathogens (porcine circovirus 2, porcine reproductive and respiratory virus, classical swine fever virus, pseudorabies virus, or porcine teschovirus). This assay detected EMCV titers at least 104 smaller than the routine PCR assay. To increase our understand of EMCV pathogenesis, the new method was used to quantify levels of EMCV genome in various tissues of artificially challenged sows and piglets. The virus was found mainly in the heart, lung, spleen, kidney, and endometrium of sows, and mainly in the heart, spleen, lung, and testis of fetuses. The real-time PCR method described here should be useful for the study of EMCV infection and distribution in pigs.

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Acknowledgments

This work was supported by National Natural Science Foundation of China (No. 31001069, No. 31172349, and No. 31172341).

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Authors and Affiliations

  1. Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, 427 Maduan Street, Nangang District, Harbin, 150001, Heilongjiang, China

    Zhao Wang, Wencheng Lin & Shangjin Cui

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

    Yebing Liu

Authors
  1. Zhao Wang
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  2. Yebing Liu
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  3. Wencheng Lin
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  4. Shangjin Cui
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Corresponding author

Correspondence to Shangjin Cui.

Additional information

Zhao Wang, Yebing Liu contributed equally to this work.

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Wang, Z., Liu, Y., Lin, W. et al. A real-time PCR to detect and analyze virulent EMCV loads in sows and piglets. Mol Biol Rep 39, 10013–10017 (2012). https://doi.org/10.1007/s11033-012-1870-y

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  • DOI: https://doi.org/10.1007/s11033-012-1870-y

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