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


CONCLUSIONS
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.
Schaeffer
Anke
A
Institute of Medical Microbiology and Hospital Hygiene, Clinical Center of Heinrich-Heine University, Moorenstrasse 5, 40225 Duesseldorf, Germany. ankeschaeffer@gmx.de
Henrich
Birgit
B
D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques; D002690:Chlamydia Infections / Q000453:epidemiology / Q000382:microbiology*; D002692:Chlamydia trachomatis / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers; D005260:Female; D006801:Humans; D008219:Lymphogranuloma Venereum / Q000453:epidemiology / Q000382:microbiology*; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D018272:Porins / Q000235:genetics; D012703:Serotyping; D013045:Species Specificity; D019914:Taq Polymerase; D013997:Time Factors
D001425:Bacterial Outer Membrane Proteins; D017931:DNA Primers; C410162:PmpH protein, Chlamydia trachomatis; D018272:Porins; C071677:omp1 protein, Chlamydia trachomatis; D019914:Taq Polymerase