An important proportion of genital samples submitted for Chlamydia trachomatis detection by PCR contain small amounts of cellular DNA as measured by beta-globin gene amplification
F. Coutlee et al., An important proportion of genital samples submitted for Chlamydia trachomatis detection by PCR contain small amounts of cellular DNA as measured by beta-globin gene amplification, J CLIN MICR, 38(7), 2000, pp. 2512-2515
We assessed the quality of genital samples submitted for Chlamydia trachoma
tis detection by PCR by a second PCR assay for the presence of human beta-g
lobin DNA, Endocervical and urethral samples were first tested by the COBAS
AMPLICOR C. trachomatis assay (Roche Diagnostic Systems) with an internal
control and were then amplified for the presence of beta-globin DNA with pr
imers PC04 and GH20. Samples that contained inhibitors were retested after
dilution 1:10. A total of 407 genital samples (311 endocervical swabs from
311 women and 96 urethral swabs from 95 men and 1 woman) collected over a 1
-month period were evaluated. The internal control could not be amplified,
despite dilution, from 3 of 23 samples that were retested after dilution be
cause of inhibition, leaving 404 samples that could be analyzed by PCR. Ele
ven samples tested positive for C, trachomatis. Thirty (7.4%) of the 404 sa
mples were negative for beta-globin. Twelve of the 23 undiluted samples tha
t contained inhibitors tested positive for beta-globin DNA. Amplification o
f beta-globin DNA in samples submitted for C, trachomatis detection by the
COBAS AMPLICOR C, trachomatis assay demonstrated that an important proporti
on of the samples did not contain cellular DNA. Assessment of the quality o
f the samples for PCR analysis by beta-globin amplification is feasible but
cannot replace use of the internal control.