title: Assessment of Chlamydia trachomatis infection by Cobas Amplicor PCR and in-house LightCycler assays using PreservCyt and 2-SP media in voluntary legal abortions.
abstract: Chlamydial infection of the upper genital tract after abortion is well recognized, but routine screening for infection before termination is rare, and few centres are aware of the prevalence of post-abortion complications in their patient population. Knowledge of the patient population is the best guide for developing screening strategies. The aim of this study was to determine the prevalence of chlamydial infection in patients presenting for legal termination of pregnancy, and to assess the presence of Chlamydia trachomatis by PCR on specimens collected in either PreservCyt (ThinPrep) or 2-sucrose phosphate (2-SP) transport medium. Two hundred and eleven single, sexually active women, aged 15-26 years, attending the Gynaecology and Obstetric Hospital, Amiens, France, for surgical termination of pregnancy were enrolled in this study from June 2002 to June 2003. C. trachomatis detection using a Cobas Amplicor PCR test (Roche Diagnostics) targeting a 207 bp segment of the common cryptic plasmid and a quantitative LightCycler real-time PCR (LC-PCR) (Roche Diagnostics) targeting a 123 bp fragment within the highly conserved constant domain 3 of the single-chromosome-copy ompA gene were performed on endocervical swabs in 2-SP, and on specimens collected using a cytobrush and placed in PreservCyt medium. The in-house LC-PCR was used as a chromosomal diagnosis method and to determine the load of C. trachomatis. This method was able to detect the mutant Swedish variant with a deletion of 377 bp in the target area in the cryptic plasmid, which is the region targeted by the Cobas Amplicor PCR test. C. trachomatis was detected in 19/211 patients (9 %) by both PCR methods. Among the 19 infected women, C. trachomatis was detected by the Cobas Amplicor PCR in 16 specimens in PreservCyt (7.6 %) and in 12 endocervical swabs in 2-SP (5.7 %). Specimens from only nine women were PCR-positive in both PreservCyt and 2-SP media by this method. Cobas Amplicor PCR revealed that 10.9 and 2.3 % of the PreservCyt and 2-SP samples, respectively, contained inhibitors. The same 19 infected women were LC-PCR positive in both PreservCyt and 2-SP samples. No additional infected women were found by this last method; thus, it was concluded that none of the samples contained the new variant of C. trachomatis. The load in each sample varied from 10(2) to 10(7) copies ml(-1).
authors.affiliation: Laboratoire d'Anatomo-Pathologie, Centre Hospitalo-Universitaire d'Amiens, Amiens, France.
authors.forename: Henri
authors.initials: H
authors.lastname: Sevestre
authors.affiliation: Centre de Gynécologie-Obstétrique, Centre Hospitalo-Universitaire d'Amiens, Amiens, France.
authors.forename: Jacques
authors.initials: J
authors.lastname: Mention
authors.affiliation: Centre de Ressources Régionales en Biologie Moléculaire, Université de Picardie Jules Verne, Amiens, France.
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Lefebvre
authors.affiliation: Service de Bactériologie, Centre Hospitalo-Universitaire d'Amiens, Amiens, France.
authors.forename: François
authors.initials: F
authors.lastname: Eb
authors.affiliation: Service de Bactériologie, Centre Hospitalo-Universitaire d'Amiens, Amiens, France.
authors.forename: Farida
authors.initials: F
authors.lastname: Hamdad
mesh_terms: D000029:Abortion, Legal; D000293:Adolescent; D000328:Adult; D001331:Automation / Q000379:methods; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D002690:Chlamydia Infections / Q000175:diagnosis* / Q000382:microbiology; D002692:Chlamydia trachomatis / Q000302:isolation & purification*; D003470:Culture Media; D005260:Female; D006801:Humans; D016133:Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods; D011247:Pregnancy; D013048:Specimen Handling / Q000379:methods
chemical_list: D001425:Bacterial Outer Membrane Proteins; D003470:Culture Media; C078167:OMPA outer membrane proteins