Resumo

JURADO-OREJUELA, Diana; PAREDES-AMAYA, Claudia  e  LIBREROS-ZUNIGA, Gerardo. Technical validation of a polymerase chain reaction for Chlamydia trachomatis detection. Salud, Barranquilla [online]. 2016, vol.32, n.3, pp.398-410. ISSN 0120-5552.

Abstract Objective: To standardize a PCR for Chlamydia trachomatis detection. Materials and methods: An experimental study was designed to standardize a C. trachomatis PCR test. Genomic DNA from C. trachomatis serovar D ATCC VR885D was used for the PCR standardization. An amplicon of 201 bp from clamidial plasmid was obtained using primers CtP15'-TAGTAACTGCCACTTCATCA-3' and CtP2 5'- TTCCCCTTGTAATTCGTT-GC-3'. Serial dilutions of clamidial DNA were used to determine the analytical sensitivity. Analytical specificity was tested using DNA from several urogenital microorganisms. Intra assay variability was assessed on triplicate DNA samples, while inter assay variability was assessed comparing the results by three technicians on different days. Results: Established (94°C/4 min; 40 cycles at 94 °C/1 min, 56°C/1 min and 72°C/1.5 min; 72°C/4 min; 1.5 mM of MgCl₂ and 1U/pL of Taq polymerase. The analytical sensitivity was 10^-17 g of DNA equivalent to one plasmid or less than one elementary body from C. trachomatis. Primers CtP1 and CtP2 amplified specifically C. trachomatis in the experimental conditions evaluated. Intra and inter assay variability demonstrated the repeatability and reproducibility of the PCR respectively. Conclusions: We standardized the experimental conditions for a C. trachomatis PCR that can be used for diagnostic purposes. Other studies are required for further clinical evaluation of this test.

Palavras-chave : Chlamydia trachomatis; molecular diagnosis; PCR.

        · resumo em Espanhol     · texto em Espanhol     · Espanhol ( pdf )