Early detection of Chlamydia trachomatis using fluorescent, DNA binding dyes.

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Abstract
HeLa 229 [human cervical carcinoma] cells were infected with genital tract strains of C. trachomatis. After incubation for varying times the infected cells were fixed and stained with the fluorescent DNA binding dyes Hoechst 33258 or DAPI [4''-6-diamidino-2-phenylindole hydrochloride] for comparison with conventional Giemsa stain. Fluorochrome-treated preparations were examined by incident UV fluorescence microscopy and the Giemsa-stained preparations by dark-ground light microscopy. Chlamydial inclusion bodies could be identified unambiguously 18 h after infection of HeLa 229 cells using Hoechst 33258 or DAPI but not until some 48 h in Giemsa-stained preparations. The DNA-rich chlamydial elementary bodies in infected egg yolk suspension were readily detected using Hoechst 33258. The fluorescent dye technique was simpler and more rapid than Giemsa staining. Hoechst 33258 can speed up the identification of chlamydial isolates growing in tissue culture.