Use of a commercial PCR kit for detecting Chlamydia trachomatis.

Smith, I. P.; Morrison, Clive L.; Patrizio, C; McMillan, Alison

doi:10.1136/jcp.46.9.822

Cited by 17 publications

(18 citation statements)

References 19 publications

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“…A number of previous studies have compared various commercial assays, culture systems, in house PCRs, and IF for C trachomatis, [10][11][12] although few have evaluated more than two assays using a common set of specimens. This study was designed to carry out a comprehensive analysis of currently available diagnostic assays and used a variety of criteria to define positive samples in the calculation of the sensitivity and specificity of each.…”

Section: Discussionmentioning

confidence: 99%

Detection of Chlamydia trachomatis in genital swabs: comparison of commercial and in house amplification methods with culture.

et al. 1998

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Aims:To evaluate the sensitivity of the Roche Cobas, Roche Amplicor plate kit, ligase chain reaction (LCR), and an in house polymerase chain reaction (PCR) by titration of purified elementary bodies (EB) and also to test 245 urethral and endocervical specimens for Chlamydia trachomatis by the four assays as well as conventional culture. Study design: EB titrations were run in duplicate in each commercial assay and six times in the in house PCR. Clinical samples were aliquoted and tested by each assay and were considered positive if C trachomatis was detected by two or more separate tests or if the sample was either culture or immunofluorescence positive. Major outer membrane protein (MOMP) specific primers were used as a confirmatory assay for the in house PCR. Results: The in house PCR, Roche Cobas Amplicor, LCR, and Amplicor plate kit gave detection limits of approximately 1, 1-2, 2, and 2-4 EBs respectively. By the criteria described above for definition of a C trachomatis positive result in clinical samples we identified 23 true positives among the 245 clinical specimens. The in house PCR detected all 23 giving a sensitivity of 100% and a specificity of 98%. The Roche Cobas Amplicor, Roche Amplicor plate kit, and LCR detected 21, 19, and 19 of these respectively giving sensitivities of 87.5%, 82%, and 82% respectively and specificities of 99.5%, 99%, and 100% respectively. The culture gave a sensitivity of 78% and specificity of 100%. Conclusion: All four amplification assays had a greater sensitivity than the culture used routinely in this laboratory. The in house plasmid PCR had the greatest sensitivity and when combined with confirmation by immunofluorescence detected the greatest number of positives. This increased sensitivity is likely to have been achieved by the use of a DNA purification step and of nested primers in the amplification stage and their combined use in routine diagnostic assays for chlamydia might increase the frequency of C trachomatis detections. However, this assay is much less user friendly than the two semiautomated commercial assays investigated in this study. (Sex Transm Inf 1998;74:289-293)

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Section: Discussionmentioning

confidence: 99%

Detection of Chlamydia trachomatis in genital swabs: comparison of commercial and in house amplification methods with culture.

et al. 1998

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“…A commercial PCR kit (Amplicor; Hoffmann-La Roche, Basel, Switzerland) was used to detect the microorganism in the cervical and endometrial specimens [7][8][9]. All the PCR analyses took place at the same laboratory and were carried out by the same biologist.…”

Section: Methodsmentioning

confidence: 99%

Detection of Chronic Endometritis by Diagnostic Hysteroscopy in Asymptomatic Infertile Patients

Polisseni

Bambirra²,

Camargos³

2003

Gynecol Obstet Invest

101

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Chronic endometritis has been related to infertility and recurrent abortion. It is usually asymptomatic, and the diagnosis is rarely clinically suspected. We performed a prospective study to evaluate both the role of diagnostic hysteroscopy in the detection of chronic endometritis in infertile patients and Chlamydia trachomatis is a potential etiologic factor. Fifty consecutive patients who sought treatment for infertility in a tertiary academic hospital were submitted to diagnostic hysteroscopy and an endometrial biopsy for histopathological study and for diagnosis of C. trachomatis by polymerase chain reaction. The patients’ mean age was 33.7 ± (SD) 5.4 years, and the duration of the couples’ infertility ranged from 1 to 18 years. The overall prevalence of chronic endometritis was 12% (6 patients). Among all patients, no cases of chlamydial infection were detected by polymerase chain reaction. In the detection of chronic endometritis, with 95% confidence intervals, the hysteroscopy sensitivity was 16.7% (range 0.9–63,5%), the specificity was 93.2% (range 80.3–98.2%), the positive predictive value was 25% (range 1.3–78.1%), and the negative predictive value was 89.1% (range 65.6–95.9%). These data suggest that hysteroscopy is not useful in the screening for chronic endometritis in asymptomatic infertile women. Further studies are needed to establish the etiology of endometritis in infertile patients.

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“…Ideally, the sensitivity of tissue culture should be 90 percent or better, but sensitivity is highly dependent on the degree to which strict requirements for transport and storage are maintained (7,11,13). Moreover, negative cultures in clearly symptomatic individuals positive for Chlamydia using other techniques have been reported (7,11).…”

Section: Diagnostic Tests For C Trachomatismentioning

confidence: 99%

“…A number of in-house polymerase chain reaction tests have been used for detecting C. trachomatis using labeled DNA probes that detect specific nucleotide sequences to ribosomal RNA, and commercial kits are available. The tests are highly sensitive and specific for C. trachomatis, ranging in sensitivity from 98 to 100 percent with 99-100 percent specificity (13,14,16). Chlamydial PCR tests were evaluated in trachoma endemic regions of Tanzania and The Gambia (14,16).…”

Section: Diagnostic Tests For C Trachomatismentioning

confidence: 99%