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Genotyping of Pathogenic Serotypes of S. suis with Pyrosequencing

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Advances and Clinical Practice in Pyrosequencing

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Abstract

Streptococcus suis (S. suis for short) can cause a variety of infections in pigs, and the infections have brought about great losses in the swine industry and some cases of deaths in human beings. In order to rapidly diagnose and control the infections of S. suis, we designed a pyrosequencing-based assay to identify the serotypes of S. suis. In the assay, pyrosequencing is used to genotype most of the pathogenic serotypes of S. suis by detecting five informative regions on the Chaperonin 60 (cpn60) gene and one species-specific region on the 16S rRNA gene, and further a few undistinguished serotypes by pyrosequencing were finely discriminated by multiplex PCR of serotype-specific fragments on the cps gene as well as species-specific fragments on the 16S rRNA gene. Through carefully designing the dispensing order of dNTP for each pyrosequencing reaction, the serotypes of S. suis could be discriminated by four pyrosequencing reactions within 3 h. Five reference serotypes and three clinical strains were successfully detected and genotyped by our assay. The results indicated that our assay is a reliable, information-rich diagnostic method for the accurate detection of S. suis serotypes.

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Author information

Authors and Affiliations

  1. Department of Pharmaceutical Analysis, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing, 210009, Jiangsu, China

    Huiyong Yang & Huan Huang

  2. Huadong Research Institute for Medicine and Biotechnics, Nanjing, China

    Haiping Wu

  3. Department of Pharmacology, Jinling Hospital, Medical School of Nanjing University, No. 305, Zhongshan East Road, Nanjing, 210002, China

    Bingjie Zou & Guohua Zhou

  4. Central Research Laboratory, Hitachi, Ltd., Tokyo, Japan

    Hideki Kambara

  5. Key Laboratory of Drug Quality Control and Pharmacovigilance, Ministry of Education, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing, Jiangsu, 210009, China

    Qinxin Song

Authors
  1. Huiyong Yang
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  2. Huan Huang
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  3. Haiping Wu
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  4. Bingjie Zou
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  5. Qinxin Song
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  6. Guohua Zhou
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  7. Hideki Kambara
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Corresponding authors

Correspondence to Qinxin Song or Guohua Zhou .

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Editors and Affiliations

  1. State Key Laboratory of Analytical Chemi, Medical School of Nanjing University, Nanjing, Jiangsu, China

    Guohua Zhou

  2. Key Laboratory of Drug Quality Control, China Pharmaceutical University, Nanjing, Jiangsu, China

    Qinxin Song

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Yang, H. et al. (2016). Genotyping of Pathogenic Serotypes of S. suis with Pyrosequencing. In: Zhou, G., Song, Q. (eds) Advances and Clinical Practice in Pyrosequencing. Springer Protocols Handbooks. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3308-2_30

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  • DOI: https://doi.org/10.1007/978-1-4939-3308-2_30

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3306-8

  • Online ISBN: 978-1-4939-3308-2

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