Evaluation of the NucliSens basic kit for detection of Chlamydia trcachomatis and Neisseria gonorrhoeae in genital tract specimens using nucleic acidsequence-based amplification of 16S rRNA

We evaluated a new RNA amplification and detection kit, the NucliSens Basic Kit (Organon Teknika), for the detection of Chlamydia trachomatis and Neis seria gonorrhoeae in genitourinary specimens. The Basic Kit provides an ope n platform for RNA amplification and detection and contains isolation reage nts for nucleic acid extraction, nucleic acid sequence-based amplification (NASBA) reagents (enzymes and buffers), and a generic ruthenium-labeled pro be for electrochemiluminescent (ECL) detection of amplified product. Using freshly purified and titrated stocks of C. trachomatis and N. gonorrhoeae a nd in vitro-generated RNA transcripts for sensitivity determinations, the B asic Kit detected 1 inclusion-forming unit of C. trachomatis, I CFU of N. g onorrhoeae, and 100 RNA molecules of 16S rRNA for both bacteria. The clinic al performance of the Basic Kit was evaluated by testing a total of 250 spe cimens for N. gonorrhoeae by culture and NASBA and a total of 96 specimens for C. trachomatis by PCR and NASBA, The Basic Kit detected 139 of 142 N. g onorrhoeae culture-positive specimens and gave a negative result for 73 of 74 culture-negative specimens, for a sensitivity and specificity of 97.9 an d 98.7%, respectively. For C. trachomatis, the Basic Kit detected 24 of 24 PCR-positive specimens and gave a negative result for 71 of 72 PCR-negative specimens, for a sensitivity and specificity of 100 and 98.6%, respectivel y. The Basic Kit also detected specimens containing both N. gonorrhoeae and C. trachomatis, using a multiplex NASBA assay using primers for both bacte ria. The NucliSens Basic Kit offers a versatile platform for the developmen t of sensitive RNA detection assays for sexually transmitted diseases.