OBJECTIVE: Urethritis is not always caused by a single pathogen, and isolation of more than two pathogens from one patient is not uncommon. We developed a method to simultaneously detect 16 pathogens related to urethritis. METHODS: We designed specific primers used for amplification of urethritis pathogens in our 16-well microplate assay. Sixteen microliters of each reaction mixture containing template DNA was added to each well to amplify 16 pathogens simultaneously. RESULTS: After we evaluated the specificity and sensitivity of this microplate polymerase chain reaction method, we used it to detect pathogens in clinical samples. Of 163 clinical samples, 49.7% (81/163) were positive for specific pathogens, and 6.7% (11/163) showed mixed infection. A specific pathogen was not identified in 43.6% (71/163) of cases. CONCLUSIONS: We developed a 16-well microplate assay with 16 specific primers to identify pathogens associated with urethritis.
OBJECTIVE:Urethritis is not always caused by a single pathogen, and isolation of more than two pathogens from one patient is not uncommon. We developed a method to simultaneously detect 16 pathogens related to urethritis. METHODS: We designed specific primers used for amplification of urethritis pathogens in our 16-well microplate assay. Sixteen microliters of each reaction mixture containing template DNA was added to each well to amplify 16 pathogens simultaneously. RESULTS: After we evaluated the specificity and sensitivity of this microplate polymerase chain reaction method, we used it to detect pathogens in clinical samples. Of 163 clinical samples, 49.7% (81/163) were positive for specific pathogens, and 6.7% (11/163) showed mixed infection. A specific pathogen was not identified in 43.6% (71/163) of cases. CONCLUSIONS: We developed a 16-well microplate assay with 16 specific primers to identify pathogens associated with urethritis.
Authors: Jimmy Twin; Nicole Taylor; Suzanne M Garland; Jane S Hocking; Jennifer Walker; Catriona S Bradshaw; Christopher K Fairley; Sepehr N Tabrizi Journal: J Clin Microbiol Date: 2011-01-05 Impact factor: 5.948