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Mol Cell Probes. 2018 Dec 5. pii: S0890-8508(18)30183-X. doi: 10.1016/j.mcp.2018.12.002. [Epub ahead of print]

Application of random amplified polymorphism DNA and 16S-23S rDNA intergenic spacer polymerase chain reaction-restriction fragment length polymorphism to predict major Streptococcus suis clonal complexes isolated from humans and pigs.

Author information

1
Faculty of Public Health, Kasetsart University Chalermphrakiat Sakon Nakhon Province Campus, Sakon Nakhon, Thailand.
2
Faculty of Veterinary Technology, Kasetsart University, Bangkok, Thailand.
3
Faculty of Science and Engineering, Kasetsart University Chalermphrakiat Sakon Nakhon Province Campus, Sakon Nakhon, Thailand.
4
Faculty of Veterinary Medicine, University of Montreal, Quebec, Canada.
5
Faculty of Public Health, Kasetsart University Chalermphrakiat Sakon Nakhon Province Campus, Sakon Nakhon, Thailand. Electronic address: Anusak.ke@ku.th.

Abstract

Random amplification of polymorphic DNA (RAPD) and 16S-23S rDNA intergenic spacer polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were applied and evaluated to determine clonal complexes (CCs) of 684 Streptococcus suis isolates from pigs and humans. RAPD better distinguished major S. suis CCs than the PCR-RFLP method. The assay was capable of simultaneously distinguishing CC1, CC16, CC25, CC28, CC104, CC221/234, and CC233/379. PCR-RFLP could not clearly differentiate among most CCs in this study except CC16. DNA sequencing using the 16S-23S rDNA intergenic spacer distinguished between four clusters: 1) consisting of CC25, CC28, CC104, and CC233/379; 2) consisting of CC221/234; 3) consisting of CC16 (ST16); and 4) consisting of CC1. This study revealed that RAPD had a greater discriminatory power than PCR-RFLP. This assay will be useful for screening or predicting major CCs relevant to human and pig S. suis clinical isolates and for low-cost screening of large numbers of isolates with rapid analytical capacity and could be utilized in most laboratories.

KEYWORDS:

Multilocus sequence typing (MLST); Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP); Random amplification of polymorphic DNA (RAPD); Serotype; Streptococcus suis

PMID:
30528897
DOI:
10.1016/j.mcp.2018.12.002

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