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Læknablaðið - 15.07.1995, Side 30

Læknablaðið - 15.07.1995, Side 30
542 LÆKNABLAÐIÐ 1995; 81 market for some time and the QuickVue test which has been recently introduced. Materials and methods Patient population and specimen collection: The patients were high risk females attending the State Clinic for Sexually Transmitted Dis- eases (STD) in Reykjavík Iceland. Patients were enrolled in the study if they had symp- toms or were contacts of patients, known to be infected with Chlamydia trachomatis. Patients were excluded if they had received antibiotics during the two weeks prior to specimen collec- tion. Four cervical specimens were collected for the detection of Chlamydia. First, three specimens were collected for culture, Quick- Vue and the Kodak SureCell in an alternating sequence. Then a fourth specimen was collect- ed for Amplicor® PCR. The sample intended for culture was collected on a cotton swab (Medical wire Co) and put in 1.0 ml of 0.2 M sucrose phosphate buffer, antibiotics and 10% foetal calf serunr and cooled with ice. The sam- ples for QuickVue and SureCell were collected on solid shaft dacron swabs. The specimen for PCR was collected with collection kits supplied by the manufacturer. Specimens were deliver- ed to the laboratory within three hours. Speci- mens for QuickVue and SureCell were proc- essed upon arrival to the laboratory. Speci- rnens for culture and PCR were either processed upon arrival to the laboratory or frozen at -70°C until processing. Test procedures: Specimens for culture of C. trachomatis were agitated with glass beads and 0.6 ml of the buffer was added to two tubes with a monolayer of McCoy cells. The cells were centrifuged for one hour at 5000 g at 35°C. The supernatant was aspirated and re- placed with maintenance media containing cy- cloheximide. The tubes were incubated at 35°C for 48-72 hours and the slide from one of the tubes was examined stained with Fluorescent Antibody (Syva MicroTrak FA). If the slide was conclusively positive or negative the slide from the second tube was stained with iodine. If the examination of the first tube was in- conclusive the second was subcultured and the procedure repeated. The number of inclusions was recorded. The QuickVue and the SureCell tests were performed according to the manu- facturers recommendations in the package in- serts. The Amplicor® assay was performed on a Perkin Elmer thermocycler. The test was per- formed according to manufacturer’s instruc- tions. Discrepant results were resolved at the Indiana School of Medicine, Infectious Dis- ease Laboratory, by doing Amplicor® PCR and DFA (Syva MicroTrak) on the leftover culture transport medium. A patient was con- sidered infected if culture was positive or if the rapid tests and a confirmatory test were posi- tive. Results Results from 240 high risk female patients were evaluated in the study. The patients were 13 to 54 years of age with the median age of 20 (figure). The infected patients were 15 to 31

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