Rapid detection of Chlamydia trachomatis and typing of the Lymphogranuloma venereum associated L-Serovars by TaqMan PCR

BMC Infect Dis. 2008 Apr 30;8:56. doi: 10.1186/1471-2334-8-56.

Abstract

Background: Infection due to Chlamydia trachomatis is the most common sexually transmitted bacterial disease of global health significance, and especially the L-serovars causing lymphogranuloma venereum are increasingly being found in Europe in men who have sex with men.

Results: The design and evaluation of a rapid, multiplex, real-time PCR targeting the major outer membrane protein (omp-1) -gene and a L-serovar-specific region of the polymorphic protein H (pmp-H) -gene for the detection of Chlamydia trachomatis is reported here. The PCR takes place as a single reaction with an internal control. For L1-, L2- and L3-serovar differentiation a second set of real-time PCRs was evaluated based on the amplification of serovar-specific omp-1-regions. The detection limit of each real-time PCR, multiplexed or not, was 50 genome copies per reaction with an efficiency ranging from 90,5-95,2%. In a retrospective analysis of 50 ocular, rectal and urogenital specimens formerly tested to be positive for C. trachomatis we identified six L2-serovars in rectal specimens of HIV-positive men, one in a double-infection with L3, and one L2 in a urethral specimen of an HIV-negative male.

Conclusion: This unique real-time PCR is specific and convenient for the rapid routine-diagnostic detection of lymphogranuloma venereum-associated L-serovars and enables the subsequent differentiation of L1, L2 and L3 for epidemiologic studies.

Publication types

  • Evaluation Study

MeSH terms

  • Bacterial Outer Membrane Proteins / genetics
  • Bacterial Typing Techniques
  • Chlamydia Infections / epidemiology
  • Chlamydia Infections / microbiology*
  • Chlamydia trachomatis / classification*
  • Chlamydia trachomatis / genetics
  • Chlamydia trachomatis / isolation & purification*
  • DNA Primers
  • Female
  • Humans
  • Lymphogranuloma Venereum / epidemiology
  • Lymphogranuloma Venereum / microbiology*
  • Male
  • Polymerase Chain Reaction / methods*
  • Porins / genetics
  • Serotyping
  • Species Specificity
  • Taq Polymerase*
  • Time Factors

Substances

  • Bacterial Outer Membrane Proteins
  • DNA Primers
  • PmpH protein, Chlamydia trachomatis
  • Porins
  • omp1 protein, Chlamydia trachomatis
  • Taq Polymerase