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.

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.