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At least 127 records · Page 7Linked to original sources
PubMed1999

Multicenter evaluation of the fully automated COBAS AMPLICOR PCR test for detection of Chlamydia trachomatis in urogenital specimens.

The fully automated COBAS AMPLICOR CT/NG test for the detection of Chlamydia trachomatis was evaluated in a multicenter trial. Test performance was evaluated for 2,014 endocervical swab and 1,278 urine specimens obtained from women and for 373 urethral swab and 254 urine specimens obtained from men. Culture served as the reference test. Culture-negative, COBAS AMPLICOR-positive specimens that tested positive in a confirmatory PCR test for an alternative target sequence within the C. trachomatis major outer membrane protein gene were resolved as true positives. The overall prevalence of chlamydia was 4.3% in cervical swabs and 11.0% in urethral swabs from men. When the results for each specimen type were considered separately, the resolved sensitivities were 96.5% (83 of 86) for endocervical swab specimens, 95.1% (39 of 41) for urine specimens from women, 100.0% (41 of 41) for urethral swab specimens from men, and 94.4% (17 of 18) for urine specimens from men; the resolved specificities were 99.4% (1,912 of 1,924) for endocervical swab specimens, 99.8% (1,204 of 1,207) for urine specimens from women, 98. 5% (325 of 330) for urethral swab specimens from men, and 100.0% (236 of 236) for urine specimens from men. For the subset of patients from whom both swab and urine specimens were collected, the combined results for both specimen types were used to identify all infected patients. Using these combined reslts as criteria, the resolved sensitivities for the COBAS AMPLICOR test were 82.6% (38 of 46) for endocervical swab specimens, 84.4% (38 of 45) for urine specimens from women, 84.2% (16 of 19) for urethral swab specimens from men, and 89.5% (17 of 19) for urine specimens from men. In comparison, the sensitivity of culture was only 56.5% (26 of 46) for endocervical specimens and 63.2% (12 of 19) for urethral specimens from men. The internal control provided in the COBAS AMPLICOR test revealed that 2.9% of specimens were inhibitory when they were initially tested. Nevertheless, valid results were obtained for 99. 1% of specimens because 68.7% of the inhibitory specimens were not inhibitory when a second aliquot of the original sample was tested. Two additional COBAS AMPLICOR-positive specimens were detected by retesting inhibitory specimens. The COBAS AMPLICOR CT/NG test for the detection of C. trachomatis exhibited equally high sensitivities and specificities with both urogenital swab and urine specimens and, thus, is well-suited for use in screening.

Automation↗
PubMed1996

[Evaluation of a new detection method of Mycobacterium tuberculosis in clinical specimens by amplification of ribosomal RNA and its clinical application].

A new rapid M. tuberculosis detection method, the Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test (MTD), which allows direct detection of M.tuberculosis in clinical specimens by amplification of M.tuberculosis ribosomal RNA (rRNA), was evaluated. In the present study, MTD and conventional smear and culture examinations were performed on 225 sputum, 36 pleural fluid (PF), nine cerebrospinal fluid (CSF) and 41 bronchoalveolar lavage fluid (BALF) specimens. Among 225 sputum specimens from 132 patients, 20 culture-negative specimens from 14 patients and one specimen which was culture-positive for M.fortuitum on Ogawa's egg medium were MTD-positive. On the other hand, there were 6 M.tuberculosis culture-positive but MTD-negative specimens from 6 patients. The amount of bacilli in all six cases, however, was very few. The sensitivity of M.tuberculosis detection by MTD, as compared with Ogawa's egg medium and MB-Check, was 93 and 92%, respectively, in sputum specimens, and the specificity of MTD was 80 and 85%, respectively. Taking into account not only culture-positive specimens but also MTD-positive specimens from patients clinically diagnosed as active tuberculosis, the sensitivity and specificity of MTD were 93% and 99%, respectively. Among 36 PF specimens from 31 patients, of which 20 specimens from 16 patients were clinically diagnosed as tuberculous, only 4 from 4 patients were culture-positive for M.tuberculosis while 7 from 5 patients were MTD-positive. The results of MTD in 9 CSF specimens coincided well with those of MB-Check, but one MTD-positive specimen yielded a false-negative result with Ogawa's egg medium. Among 41 BALF specimens from 39 patients, only one was culture-positive while one culture-positive and 7 culture-negative specimens were positive on MTD. All these MTD-positive patients were later verified as having pulmonary tuberculosis either by transbronchial lung biopsy and other examinations, or by the clinical course of the disease, particularly the response to anti-tuberculosis drugs. Inconsistent results were seen in 21 sputum specimens from 15 patients, 4 PF specimens from 3 patients and 7 BALF specimens from 7 patients, all of which were culture-negative for M.tuberculosis but MTD-positive. None of these patients had any clinical findings inconsistent with diagnosis as tuberculosis. It was noted that MTD was very useful for rapid detection of M.tuberculosis in sputum as well as other clinical specimens. We observed the clinical course of 20 pulmonary tuberculosis patients by monthly examinations of sputum, using smears, Ogawa's egg medium, MB-Check and MTD. MTD-positivity rates fell down in parallel with decreased pulmonary tuberculous activity. During the clinical course, out of 61 MTD-negative specimens only 4 (6.6%) were culture-positive for M.tuberculosis, while out of 63 smear-and-culture-negative specimens 6 (9.5%) were MTD-positive. These data suggest that MTD is useful to assess the clinical activity and course of tuberculosis.

Adult↗
PubMed1997

Detection of Neisseria gonorrhoeae and Chlamydia trachomatis in genitourinary specimens from men and women by a coamplification PCR assay.

A coamplification PCR test for the direct detection of Neisseria gonorrhoeae and Chlamydia trachomatis in urethral and endocervical swabs and urine samples from men and women was compared to standard culture techniques. Processed specimens were amplified in single reaction tubes containing primers for both organisms, and PCR products were detected by a colorimetric microwell plate hybridization assay specific for each pathogen. Of 344 specimens from men, 45 (13.1%) urine specimens were PCR positive for C. trachomatis, 51 (14.8%) urethral swab specimens were PCR positive, and 29 urethral swab specimens (8.4%) were culture positive. After analysis of discrepancies, the resolved sensitivity and specificity of PCR for C. trachomatis were 96.2 and 99.3%, respectively, in urethral swab specimens, compared to 88.2 and 98.6% for urine specimens. Of the 192 specimens from women, 28 (14.6%) urine specimens were PCR positive for C. trachomatis, 32 (16.7%) endocervical specimens were PCR positive, and 19 (9.9%) endocervical specimens were culture positive. After analysis of discrepancies, the resolved sensitivity and specificity of PCR for C. trachomatis for endocervical specimens were both 100% compared to 100 and 99.4%, respectively, for urine specimens from women. In men, 68 (19.8%) urine specimens were PCR positive for N. gonorrhoeae, 73 (21.2%) urethral swabs were PCR positive, and 59 (17.2%) urethral swabs were culture positive. After analysis of discrepancies, the resolved sensitivity and specificity of PCR for N. gonorrhoeae were 97.3 and 97.0%, respectively, for urethral specimens compared to 94.4 and 98.5% for urine specimens. In women, 18 (9.4%) urine specimens were PCR positive for N. gonorrhoeae, 23 (12.0%) were endocervical swab PCR positive, and 15 (7.8%) endocervical specimens were culture positive. After analysis of discrepancies, the resolved sensitivity and specificity of PCR for N. gonorrhoeae were 100 and 99.4%, respectively, for endocervical specimens compared to 90.0 and 95.9% for female urine specimens. These results indicate that a multiplex PCR is highly sensitive for detecting both C. trachomatis and N. gonorrhoeae from a single urine or genital swab, providing a more cost-effective way of screening multiple pathogens.

Chlamydia Infections↗