Loss of virulence in Shigella strains preserved in culture collections due to molecular alteration of the invasion plasmid.

Fifty-two Shigella strains long preserved by three Japanese culture collections were examined for virulence. All of them were avirulent when judged by the focus-plaque assay and the ability to bind Congo red. Fifteen strains had a plasmid comparable in size to that responsible for epithelial invasiveness and were positive in hybridization tests with a probe derived from a plasmid cistron, virG. Twenty-four strains had a similar plasmid but were negative in hybridization tests. The remaining 13 strains were negative in all the five criteria for virulence. Similar studies made on one hundred Shigella strains isolated from 1967 to 1985 clearly demonstrated loss of virulence with prolonged time of storage.