J Clin Pathol Qual Control.  1999 Dec;21(2):295-299.

Detection of Chlamydia trachomatis by Dual PCR

• Lee SY
• Oh JH
• Jang JS
• Kim DK
• Kim DA
• Choi TY

Abstract

BACKGROUND: The cell culture method is used as the gold standard for detection of C. trachomatis, but it is very laborious and time consuming. In this study, we detected C. trachomatis by dual polymerase chain reaction (PCR) and compared its diagnostic performance with those of the cell culture method.
METHODS
This study included 504 patients with pelvic inflammatory diseases or non gonococcal urethritis, or conjunctivitis. The samples were inoculated onto McCoy cell monolayers in shell vials. Two primer sets were chosen from the dual PCR ; one detected a specific sequence of the plasmid, and the other detected the omp1 gene encoding the major outer membrane proteins.
RESULTS
In control strains, both primer sets reacted specifically and amplified both the omp1 gene (493 bp) and the plasmid gene (144 bp) sequence from all human serovars simultaneously. In two of the samples, C. trachomatis was isolated using the cell culture method. The prevalence rate of C. trachomatis tested by the dual PCR was 3% (15 of the 504 samples). Twelve of the 15 positive samples showed positive responses by omp1 gene and plasmid DNA amplification. Two of 15 positive samples were found to be positive only by plasmid amplification.
CONCLUSION
On the basis of these results, it is considered that the dual PCR technique can be applied as an effective tool for diagnosing C. trachomatis infections.

MeSH Terms

Cell Culture Techniques
Chlamydia trachomatis*
Chlamydia*
Conjunctivitis
DNA
Female
Humans
Membrane Proteins
Pelvic Inflammatory Disease
Plasmids
Polymerase Chain Reaction*
Prevalence
Urethritis
DNA
Membrane Proteins