The present study describes the prevalence of muraminidase-released protein (MRP) and extracellular factor (EF) proteins associated with virulence of Streptococcus suis serotype 2 from a collection of USA strains. Sixty-six strains belonging to serotypes 1, 2, 3, 4, 7, and 10, were analyzed with a set of double antibody sandwich ELISAs and Western blots. Nineteen of 34 serotype 2 strains from cases of swine meningitis had the MRP+EF+ phenotype. Five of 7 serotype 2 strains isolated from lungs had an MRP*EF- phenotype. An MRP-EF+ phenotype was found in 4/34 strains isolated from swine meningitis. The MRP*EF- and MRP-EF+ phenotypes have not been reported previously. All strains of serotypes other than 2, including isolates from cases of meningitis, had the MRP-EF- phenotype, suggesting that these strains must have other, as yet undetected, virulence factors.