Immunomagnetic isolation of Streptococcus suis serotypes 2 and 1/2 from swine tonsils

Abstract

Isolation of specific serotypes of Streptococcus suis from the tonsils, nasal cavities, and genital tract is difficult, since low-pathogenic serotypes and untypeable strains also inhabit these sites. An immunomagnetic separation (IMS) technique for the selective isolation of S. suis serotypes 2 and 1/2 was standardized. Superparamagnetic polystyrene beads (immunomagnetic beads or IMB) were coated with either a purified monoclonal antibody (MAb) directed to a capsular sialic acid-containing epitope or purified rabbit immunoglobulin G (polyclonal antibody [PAb]), both specific for S. suis serotypes 2 and 1/2. The amount of antibodies required for optimum coating of the beads, the number of IMB required for optimum bacterial recovery, and the nonspecific carryover were considerably higher with the MAb-IMS technique than with the PAb-IMS technique. The sensitivity of the IMS technique was 10(1) CFU/0.1 g of tonsil. The presence of serotype 1/2 bacteria did not considerably affect the recovery rate of a serotype 2 strain and vice versa. To validate the technique, PAb-coated beads were used to study 192 tonsils from animals from S. suis serotype 2- or 1/2-infected herds. Results showed that significantly more positive tonsils were detected by the IMS technique than by the standard procedure. This method represents an innovative and highly sensitive approach for the isolation of S. suis serotypes 2 and 1/2 from carrier animals.

FIG. 1

Recovery of S. suis serotype 2 with an initial count of 10³ CFU (⧫) or 10⁶ CFU (×) using IMB coated with different concentrations of S. suis serotype 2- and 1/2-specific MAb Z3 (A) or PAb IgG (B).

FIG. 2

Recovery of S. suis serotype 2 (initial count of 10³ CFU) using different concentrations of IMB coated with serotype 2- and 1/2-specific MAb Z3 (⧫) or PAb IgG (×).