Detection of C trachomatis in urogenital specimens by polymerase chain reaction.
Open Access
- 1 June 1991
- journal article
- research article
- Published by BMJ in Sexually Transmitted Infections
- Vol. 67 (3), 211-214
- https://doi.org/10.1136/sti.67.3.211
Abstract
OBJECTIVE--To establish a polymerase chain reaction (PCR) protocol for the detection of urogenital C trachomatis infection and to compare it with the detection in cell culture. SPECIMENS--Urethral specimens were collected from 62 male patients and cervical specimens from 106 female patients. SETTING--Department of Dermatology and Venereology, Ruprecht-Karls-Universität, Heidelberg. METHODS--Urogenital specimens were simply boiled for 15 minutes and subsequently subjected to amplification without prior extraction of nucleic acid. The DNA sequence selected for amplification is located in the third open reading frame of the ubiquitous C trachomatis plasmid pCTT1. The amplified products were demonstrated by agarose gel electrophoresis followed by Southern blot hybridization. In addition, specimens were investigated with cell culture. MAIN OUTCOME MEASURES--Results of PCR and cell culture. RESULTS--PCR detected all C trachomatis serovars relevant for urogenital infections (D-L2). Serial dilution experiments revealed that the PCR procedure was 100 fold more sensitive than cell culture. The investigation of 168 urogenital specimens showed that the PCR confirmed all 30 cell culture positive results, however, out of the 138 cell culture negative specimens 16 were positive using the PCR. CONCLUSIONS--A substantial number of urogenital C trachomatis infections detectable by PCR may be missed by the cell culture technique.Keywords
This publication has 16 references indexed in Scilit:
- Evaluation of a radioactive rRNA:cDNA-hybridisation assay for the direct detection of Chlamydia trachomatis in urogenital specimens.Sexually Transmitted Infections, 1989
- Specific amplification of a DNA sequence common to all Chlamydia trachomatis serovars using the polymerase chain reactionResearch in Microbiology, 1989
- Evaluation of non-radioactive in situ hybridisation method to detect Chlamydia trachomatis in cell culture.Sexually Transmitted Infections, 1988
- Primer-Directed Enzymatic Amplification of DNA with a Thermostable DNA PolymeraseScience, 1988
- A common plasmid of Chlamydia trachomatisPlasmid, 1986
- Use of nucleic acid probes for the detection of sexually transmitted infectious agentsDiagnostic Microbiology and Infectious Disease, 1986
- Comparison of methods for detecting Chlamydia trachomatis.Journal of Clinical Pathology, 1986
- Analysis and Detection of Chlamydial DNAMicrobiology, 1984
- Culture-Independent Diagnosis ofChlamydia trachomatisUsing Monoclonal AntibodiesNew England Journal of Medicine, 1984
- Monoclonal Antibodies for Diagnosis of Infectious Diseases in HumansScience, 1983