Analyzing multiclonality of Staphylococcus aureus in clinical diagnostics using spa-based denaturing gradient gel electrophoresis

Abstract

We present a novel denaturing gradient gel electrophoresis (DGGE) method which characterizes multiclonal communities of Staphylococcus aureus. The spa PCR-based DGGE method simultaneously separates strains that differ in only one base, thereby revealing multiclonal colonization and infections.

Fig. 1.

spa-DGGE analysis of the eight different strains (lanes 1 to 8, t064, t355, t012, t3061, t050, t008, t002, and t015, respectively; lane 9, a mixture of equal amounts of eight PCR products).

Fig. 2.

Broth samples from the outbreak containing MRSA isolates of spa type t015 or t069 were analyzed by the spa-DGGE method. In samples from two individuals, both spa types, t015 and t069, were detected (lanes 2 and 3). In lanes 4 to 7, the samples contain spa type t015, and in lanes 8 to10, the samples contain spa type t069. In lane 1, PCR products from both spa types, t015 and t069, are mixed and analyzed as a control.