Development and clinical evaluation of a polymerase chain reaction test for detection of Chlamydia trachomatis

Abstract

A polymerase chain reaction (PCR) for the detection of Chlamydia trachomatis was developed and evaluated. Two primer-probe sets were designed; one detected a specific sequence of the plasmid, and the other detected the gene encoding the major outer membrane protein. Both sets reacted species specifically and amplified sequences from all human serovars. A simple protocol was used for sample pretreatment. The PCR was optimized by addition of tetramethylammonium chloride and bovine serum albumin. The results of the PCR with the plasmid primer-probe set were compared with those of culture and the Chlamydiazyme and Gen-Probe PACE 2 tests for urogenital specimens from 220 patients. The rates of prevalence of infection with C. trachomatis were 22.7, 16.4, 15.0, and 14.5%, respectively. The sensitivities of the Chlamydiazyme and Gen-Probe PACE 2 assays compared with culture were 66.7 and 61.1%, respectively, and their sensitivities compared with PCR were 60.0 and 60.0%, respectively. The sensitivity of culture compared with PCR was 70.0%. Forty-eight of the 50 specimens positive by PCR with the plasmid primer-probe set could be confirmed by PCR with the major outer membrane protein primer-probe set or culture. It is concluded that the PCR is the most sensitive technique for laboratory detection of C. trachomatis.

Methods of detection of Chlamydia psittaci in domesticated and wild birds

“…The use of PCR detection of human C trachomatis infections has been widely evaluated. 12 A kit amplifying plasmid DNA is available for the detection of C trachomatis. Of the PCR primer sets evaluated for the detection of human chlamydial infections, those amplifying chlamydial plasmid DNA have proven the most sensitive compared to the copy chromosomal genes encoding OMP1 or 16s rRNA.…”

Amplification of plasmid and chromosome chlamydia dna in synovial fluid of patients with reactive arthritis and undifferentiated seronegative oligoarthropathies

“…The initial heating step of each round was lengthened to 5 minutes to ensure complete DNA denaturation and increase the sensitivity. DNA detection was done after gel electrophoresis, blotting, and hybridization, again to increase sensitivity (with oligonucleotide probes, it can be increased 10-fold compared with detection after ethidium bromide staining of the gels [25,26]; with our longer probes [71-380 bp], sensitivity can be expected to increase further). In addition, this procedure yields information on the size, and thus on the specificity, of lanes 3 , 6 , and 8).…”

Rapid and sensitive detection of Mollicutes in cell culture by polymerase chain reaction

“…In this study, we used the specific primers GPO-3 and MGSO [21,25,26]. The forward primer (GPO-3) was altered by adding 2 (G & T) nucleotides in the 5′ region.…”