BACKGROUND--Infection caused by Chlamydia trachomatis is now recognised as the most prevalent sexually transmitted disease in many parts of the world. Anorectal infections caused by C. trachomatis is not uncommon. Enzyme-linked immunoassay (EIA) detects an antigen lipopolysaccharide (LPS) of C. trachomatis directly in clinical specimens. OBJECTIVE--Our aim was to compare an enzyme immunoassay, Wellcozyme Chlamydia (WZ04) with cell culture for the diagnosis of chlamydial infection of the anogenital tract. METHOD--Rectal swabs were taken from 100 prostitutes (80 females and 20 males) for chlamydia culture, WZ04 and direct immunofluorescence (DIF). In addition, endocervical specimens were obtained from the females for the above three tests. MAIN FINDINGS--All the positive rectal specimens were from females. Nine patients had a positive chlamydia culture from the rectum but negative WZ04 and DIF. Two patients had false positive results by WZ04 but negative culture and DIF. For cervical specimens, WZ04 identified 43% (3/7) of the culture positive cases. Specificity was 98.6%. WZ04 identified an additional specimen as positive which was also confirmed as positive by DIF. CONCLUSION--Our study shows that in our hands enzyme-linked immunoassays such as Wellcozyme Chlamydia are neither sensitive nor specific in detecting C. trachomatis infection of the rectum. For cervical infections, the sensitivity of WZ04 was 43% and the specificity 98.6% as compared to culture, with a positive predictive value of 75% and a negative predictive value of 94.7%.