Detection ofChlamydia trachomatisby nucleic acid sandwich hybridization

Chromosomal DNA fragments from Chlamydia trachomatis serotype L2 were shotgun-cloned into pBR322. A C. trachomatis-specific clone was further subcloned to produce specific DNA reagents for the identification of C. trachomatis by nucleic acid sandwich hybridization. The chosen DNA reagents from serotype L2 also hybridized with all the other chlamydial serotypes tested, but not with the DNA from 41 unrelated organisms. The sensitivity of the sandwich hybridization test was 106 DNA molecules. The applicability of the test for routine diagnostic use was demonstrated by a pilot study in which C. trachomatis was directly detected from genital specimens.