SUMMARY: One hundred and ninety-six urethral and endocervical swabs were processed for isolation of C. trachomatis, using H-1 HeLa cells on shell vials, in the presence and absence of 7% PEG in the chlamydial overlay medium. The results were divided into three evaluable groups based on the number of inclusions per coverslip in the shell vials without PEG. The number of inclusions were compared with that of PEG-treated cultures using the paired t-test. The ranges for the three evaluable groups were one to ten, 11–100 and 101–1000 inclusions per coverslip. All three groups showed a significant increase (2.8- to 3.8-fold) in the number of inclusion bodies in the PEG-treated cultures compared to the untreated cultures.