Abstract
The aim of this study was to assess the presence of inhibitors in urine specimens causing false-negative results in a commercialChlamydia trachomatis gap-filling ligase chain reaction (Gap-LCR) assay. On testing of urine samples by the Gap-LCR assay and urethral swab specimens by cell culture, 73 (19%)Chlamydia trachomatis positive subjects were detected among 382 men attending a clinic for sexually transmitted diseases. In 56 subjects, the agent was detected in both the urine and the urethral samples, while 309 subjects were negative in both tests. In seven subjects urine samples were Gap-LCR positive (confirmed by a different Gap-LCR assay), but the corresponding urethral swab samples were cell culture-negative. In another ten subjects the urethral swab samples were cell culture positive, but their urine samples were Gap-LCR negative. Subsequent re-analysis of the urine samples including the addition of externalChlamydia trachomatis DNA indicated full or partial inhibition in nine of the cell culture-positive Gap-LCR negative subjects. When urine preparations were freeze-thawed and diluted prior to testing,Chlamydia trachomatis was detected in six of the ten initially Gap-LCR-negative samples. Gap-LCR inhibitors were present in at least nine (12%) of the 73 urine preparations from theChlamydia trachomatis positive individuals. Identification of samples containing Gap-LCR inhibitors and subsequent processing to reduce the inhibition increased the sensitivity of the test from 86% to 95%.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Loeffelholz MJ, Lewinski CA, Silver SR, Purohit AP, Herman SA, Buonagurio DA, Dragon EA: Detection ofChlamydia trachomatis in endocervical specimens by polymerase chain reaction. Journal of Clinical Microbiology 1992, 30: 2847–2851.
Mahony JB, Luinstra KE, Sellors JW, Jang D, Chernesky MA: Confirmatory polymerase chain reaction testing forChlamydia trachomatis in first-void urine from asymptomatic and symptomatic men. Journal of Clinical Microbiology 1992, 30: 2241–2245.
Dutilh B, Bébéar C, Rodriguez P, Vekris A, Bonnet J, Garret M: Specific amplification of a DNA sequence common to allChlamydia trachomatis serovars using the polymerase chain reaction. Research in Microbiology 1989, 140:7–16.
An Q, Radcliffe G, Vassallo R, Buxton D, O'Brien WJ, Pelletier DA, Weisburg WG, Klinger JD, Olive DM: Infection with a plasmid-free variantChlamydia related toChlamydia trachomatis identified by using multiple assays for nucleic acid detection. Journal of Clinical Microbiology 1992, 30: 2814–2821.
Dille BJ, Butzen CC, Birkenmeyer LG: Amplification ofChlamydia trachomatis DNA by ligase chain reaction. Journal of Clinical Microbiology 1993, 31: 729–731.
Ripa KT, Mårdh PA: Cultivation ofChlamydia trachomatis in cycloheximide-treated McCoy cells. Journal of Clinical Microbiology 1977, 6: 328–331.
Chernesky MA, Jang D, Lee H, Burczak JD, Hu H, Sellors J, Tomazic-Allen SJ, Mahony JB: Diagnosis ofChlamydia trachomatis infections in men and women by testing first-void urine by ligase chain reaction. Journal of Clinical Microbiology 1994, 32: 2682–2685.
Ånestad G, Berdal BP, Scheel O, Mundal R, Odinsen O, Skaug K, Khalil OS, Plier P, Lee H: Screening urine samples by leukocyte esterase test and ligase chain reaction forChlamydial infections among asymptomatic men. Journal of Clinical Microbiology 1995, 33: 2483–2484.
Lee HH, Chernesky MA, Schachter J, Burczak JD, Andrews WW, Muldoon S, Leckie G, Stamm WE: Diagnosis ofChlamydia trachomatis genitourinary infection in women by ligase chain reaction assay of urine. Lancet 1995, 345:213–216.
Bassiri M, Hu HY, Domeika MA, Burczak J, Svensson L-O, Lee HH, Mårdh PA: Detection ofChlamydia trachomatis in urine specimens from women by ligase chain reaction. Journal of Clinical Microbiology 1995, 33: 898–900.
Pasternack R, Vuorinen P, Pitkäjärvi T, Koskela M, Miettinen A: Comparison of manual Amplicor PCR, Cobas Amplicor PCR, and LCx assays for detection ofChlamydia trachomatis infection in women by using urine specimens. Journal of Clinical Microbiology 1997, 35:402–405.
Van-Doornum GJJ, Buimer M, Prins M, Henquet CJM, Coutinho RA, Plier PK, Tomazic-Allen S, Hu H, Lee H: Detection ofChlamydia trachomatis infection in urine samples from men and women by ligase chain reaction. Journal of Clinical Microbiology 1995, 33: 2042–2047.
Grimprel E, Sanchez PJ, Wendel GD, Burstain JM, McCracken GH, Radolf JD, Norgard MV: Use of polymerase chain reaction and rabbit infectivity testing to detectTreponema pallidum in amniotic fluid, fetal and neonatal sera, and cerebrospinal fluid. Journal of Clinical Microbiology 1991, 29: 1711–1718.
Wiedbrauk DL, Werner JC, Drevon AM: Inhibition of PCR by aqueous and vitreous fluids. Journal of Clinical Microbiology 1995, 33: 2643–2646.
Peterson EM, Markoff BA, Schachter J, de la Maza LM: The 7.5-kb plasmid present inChlamydia trachomatis is not essential for the growth of this microorganism. Plasmid 1990, 23: 144–148.
Van-Vollenhoven P, Heyns CF, de-Beer PM, Whitaker P, van-Helden PD, Victor T: Polymerase chain reaction in the diagnosis of urinary tract tuberculosis. Urological Research 1996, 24: 107–111.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Berg, E.S., Ånestad, G., Moi, H. et al. False-negative results of a ligase chain reaction assay to detectChlamydia trachomatis due to inhibitors in urine. Eur. J. Clin. Microbiol. Infect. Dis. 16, 727–731 (1997). https://doi.org/10.1007/BF01709252
Issue Date:
DOI: https://doi.org/10.1007/BF01709252