Development of a multiplex PCR for identification of β-hemolytic streptococci relevant to human infections and serotype distribution of invasive Streptococcus agalactiae in Thailand

A multiplex polymerase chain reaction (mPCR) was developed for simultaneous detection (single reaction) of genes specific to five frequent clinically relevant β-hemolytic streptococcal species: Streptococcus pyogenes (Spy1258), Streptococcus agalactiae (cfb and cpn60), Streptococcus dysgalactiae subsp. equisimilis (16S-23S intergenic spacer) , S. equi subsp. zooepidemicus (esaA and sorD), and Streptococcus anginosus group (moaC). No cross-reaction was observed with other bacterial species. This test was validated and successfully used with 725 clinical isolates involved in pathological conditions in Thailand and collected between March 2014 and December 2015. Results showed that S. agalactiae, mainly serotype III, was the most common Streptococcus isolated from invasive diseases. This assay should be useful for laboratory identification and surveillance of human infections by these species.