Comparison of HeLa 229 and McCoy cell cultures for detection of Chlamydia trachomatis in clinical specimens

Thewessen, E. A. P. M.; Freundt, Ingrid; Rijsoort-Vos, J. H. Van; Stolz, E.; Michel, M. F.; Wagenvoort, J. H. T.

doi:10.1128/jcm.27.6.1399-1400.1989

Cited by 15 publications

(8 citation statements)

References 9 publications

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“…Cell culture. Chlamydial culture was performed on cycloheximide-treated Mc-Coy cells in microtiter plates and has been described extensively (16,17,30). Briefly, two wells per plate were each inoculated with 0.2 ml of a patient sample.…”

Section: Methodsmentioning

confidence: 99%

Comparison of three commercially available amplification assays, AMP CT, LCx, and COBAS AMPLICOR, for detection of Chlamydia trachomatis in first-void urine

Goessens¹,

Mouton²,

Meijden³

et al. 1997

J Clin Microbiol

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We compared the Gen-Probe transcription-mediated amplification assay (AMP CT), the Abbott LCx assay, and the Roche COBAS AMPLICOR assay for the detection of Chlamydia trachomatis in a mixed population in urine samples. First-void urine, urethral specimens, and cervical specimens in females were obtained from 1,000 patients (544 males and 456 females) visiting the outpatient sexually transmitted disease clinic of our hospital. The prevalence of C. trachomatis infection was 7.7% as determined by tissue culture of urethral and cervical specimens. The sensitivities of LCx, COBAS AMPLICOR, and AMP CT compared to cell culture were 79, 86, and 78%, respectively. Sensitivity and specificity were recalculated by using a new "gold standard", i.e., a sample was considered to be true positive if two or more techniques yielded positive results. Specimens positive only by cell culture or positive in only one commercial amplification technique were retested by a previously described in-house PCR. After discordance analysis the sensitivities of LCx, COBAS AMPLICOR, and AMP CT were 84, 93, and 85%, respectively. Specificity exceeded 99% for all three assays. With each method the sensitivity was lower for urine samples from females compared to urine samples from males. By application of this new gold standard, existing differences between methods are highlighted; future evaluations of new techniques should be validated against two or more amplification assays.

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Section: Methodsmentioning

confidence: 99%

Comparison of three commercially available amplification assays, AMP CT, LCx, and COBAS AMPLICOR, for detection of Chlamydia trachomatis in first-void urine

Goessens¹,

Mouton²,

Meijden³

et al. 1997

J Clin Microbiol

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show abstract

“…Cell culture. Chlamydial culture was performed with cycloheximide-treated McCoy cells in microtiter plates and has been described previously (14,15,24). Briefly, two wells per plate were each inoculated with 0.2 ml of a patient sample.…”

Section: Methodsmentioning

confidence: 99%

Detection of Chlamydia trachomatis in male and female urine specimens by using the amplified Chlamydia trachomatis test

et al. 1997

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The amplified Chlamydia trachomatis test (AMP-CT; Gen-Probe), a new diagnostic test for the detection of Chlamydia trachomatis, was evaluated with urine specimens from 1,000 patients visiting the outpatient department for sexually transmitted diseases at the University Hospital Rotterdam, Rotterdam, The Netherlands, by comparing the results to those of cell culture. From February 1996 to July 1996, urine samples for the AMP-CT test and urethral swabs for cell culture were collected from 544 men, while cervical swabs from 456 women were also taken for cell culture. Positive test results were obtained for 130 (13%) of the patients. AMP-CT test and cell culture results were discordant for 70 (7%) specimens. Analysis of the samples with discordant results was performed by an in-house PCR. After resolution of the discordant results, the sensitivity, specificity, and positive and negative predictive values of the AMP -CT test were 84.3, 98.8, 89.6, and 98%, respectively, for samples from females and 100, 99.2, 93.1, and 100%, respectively, for samples from males, while for cell culture these values were 72.5, 99.2, 92.5, and 98%, respectively, for samples from females and 57.4, 99.0, 86.1, and 95.4%, respectively, for samples from males. We conclude that the AMP-CT test is a fast and reliable test for the detection of C. trachomatis in urine specimens from females and, in particular, males.

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“…Cell culture. C. trachomatis was cultured in cycloheximide-treated McCoy cells, grown in 96-well microtiter plates as described by Thewessen et al (27). Briefly, two wells per plate were each inoculated with 0.2 ml of patient sample.…”

Section: Methodsmentioning

confidence: 99%

“…Analysis of crude extracts from clinical specimens is hampered by the limited sample volume that can be tested in PCR and the presence of inhibitors of the PCR assay (8,27,29,30). To overcome these problems, assays that use solid-phasecoated, microbe-specific antibodies for concentration and purification of microbial particles from crude clinical specimens have been developed (11,14,20,29,31).…”

mentioning

confidence: 99%

Solid-phase C1q-directed bacterial capture followed by PCR for detection of Chlamydia trachomatis in clinical specimens

et al. 1995

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An antigen capture system based on the binding of bacteria to solid-phase immobilized complement C1q followed by PCR for detection of Chlamydia trachomatis in clinical samples was developed and clinically evaluated. Comparison of C1q-directed antigen capture PCR with cell culture and direct PCR on 71 consecutive clinical specimens revealed an identical sensitivity. In this group, all 11 cell culture-positive samples were positive by direct PCR and C1q-directed antigen capture PCR. In addition, two samples found negative by cell culture were found positive by both direct PCR and C1q-directed antigen capture PCR. To further assess the sensitivity of C1q-directed antigen capture PCR, 20 clinical samples with one to five inclusions in cell culture and 20 clinical samples with 6 to 20 inclusions in cell culture were tested. Results obtained showed sensitivities of 95 and 90% for clinical samples with 6 to 20 and 1 to 5 inclusions in cell culture, respectively. Using C1q-coated solid phases, C1q-binding Chlamydia particles can be concentrated from large volumes with concomitant removal of inhibitors of PCR, allowing the use of large volumes of clinical samples for clinical testing. Since C1q has been shown to bind to a range of gram-negative bacteria, the newly developed technique has utility for a broad range of bacteria.

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Comparison of HeLa 229 and McCoy cell cultures for detection of Chlamydia trachomatis in clinical specimens

Cited by 15 publications

References 9 publications

Comparison of three commercially available amplification assays, AMP CT, LCx, and COBAS AMPLICOR, for detection of Chlamydia trachomatis in first-void urine

Comparison of three commercially available amplification assays, AMP CT, LCx, and COBAS AMPLICOR, for detection of Chlamydia trachomatis in first-void urine

Detection of Chlamydia trachomatis in male and female urine specimens by using the amplified Chlamydia trachomatis test

Solid-phase C1q-directed bacterial capture followed by PCR for detection of Chlamydia trachomatis in clinical specimens

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