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Development of multiplex TaqMan qPCR for simultaneous detection and differentiation of eight common swine viral and bacterial pathogens

  • Veterinary Microbiology - Research Paper
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Brazilian Journal of Microbiology Aims and scope Submit manuscript

Abstract  

It is laborious to diagnose the infections of classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), and Suid herpesvirus 1 (SuHV-1) because of the similar clinical symptoms in piglets. Staphylococcus aureus (S. aureus), Streptococcus suis (S. suis), Salmonella choleraesuis (S. choleraesuis, serotype: 6,7:c:1,5), and Escherichia coli (E. coli) are common secondary bacterial pathogens in viral infections. Furthermore, the mixed infection of these viral and bacterial pathogens is more and more common in practical swine breeding. Therefore, a TaqMan multiplex qPCR method for simultaneous detection and differentiation of their pathogen was established in this study by designing specific primers and probes for the E2 gene of CSFV, the ORF7 gene of PRRSV, the ORF1 gene of PCV2 and the gE gene of SuHV-1, the nuc gene of S. aureus, the ef-tu gene of S. suis, the ivnA gene of S. choleraesuis, and the 23S rRNA gene of E. coli, and its specificity, sensitivity, and reproducibility were subsequently tested. The results showed that TaqMan multiplex qPCR method showed a high specificity with no cross reaction between different viruses, and a good repeatability with its coefficient of variation lower than 5%. Besides, the sensitivity of this method was also at least 10 times higher compared with conventional PCR. Overall, this study provided a reliable multiplex TaqMan qPCR method for the diagnosis and differentiation of the mentioned pathogens in pigs, laying a certain technical basis for disease prevention and control.

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Funding

This work was supported by grants from the science and technology projects of Yangling demonstration area (No. 2018NY-10) and the innovation project for agro-technology of Shaanxi Province (No. 2019NY-081), China.

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Authors and Affiliations

  1. College of Veterinary Medicine, Northwest A&F University, Yangling, 712100, Shaanxi, China

    Qi Zhang, Feng Yang, Jie Gao, Weimin Zhang & Xingang Xu

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  1. Qi Zhang
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  2. Feng Yang
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  3. Jie Gao
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  4. Weimin Zhang
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  5. Xingang Xu
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Contributions

Qi Zhang, Feng Yang, and Jie Gao performed the majority of experiments and involved in manuscript preparation. Xingang Xu and Weimin Zhang conceived of the study, participate in its design and coordination, and revised the manuscript. All authors read and approved the final manuscript.

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Correspondence to Weimin Zhang or Xingang Xu.

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The authors declare no competing interests.

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Qi Zhang, Feng Yang and Jie Gao are contributed equally to this work.

Responsible Editor: Fernando R. Spilki

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Zhang, Q., Yang, F., Gao, J. et al. Development of multiplex TaqMan qPCR for simultaneous detection and differentiation of eight common swine viral and bacterial pathogens. Braz J Microbiol 53, 359–368 (2022). https://doi.org/10.1007/s42770-021-00633-w

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  • DOI: https://doi.org/10.1007/s42770-021-00633-w

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