Abstract

We have reported a method for detection of Chlamydia trachomatis by polymerase chain reaction (PCR) with two oligonucleotides based on sequences within the major outer membrane protein gene from C. trachomatis serovar L2. In the previous report, in addition to treatment of the mixture of first-voided urine (FVU) sediment and 1 ml of urine with proteinase K. DNA purification by phenol extraction was necessary for preparation of template DNA for PCR. In this study, FVU sediment was suspended in 1 ml of Chlamydiazyme dilution buffer and a part of the suspension was treated with proteinase K for DNA extraction. The DNA extraction solution could be used as template for PCR without purification of DNA by phenol extraction. One hundred FVU specimens obtained from male urethritis patients were examined with the two methods (PCR and IDEIA) for detection of C. trachomatis. In 33 of 100 specimens, the DNA fragments of C. trachomatis was amplified by the PCR and in 32 of 100, the chlamydial antigen was detected by IDEIA. The positive and negative coincidence rate of the PCR to IDEIA were 93.8% (30.32) and 95.6% (65/68) respectively, resulting in a high overall coincidence rate at 95%. Thus, the improved method with PCR using FVU as a specimen is proved to be a useful, non-invasive diagnostic tool for diagnosis of chlamydial urethritis.

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