DETECTION OF Chlamydia trachomatis IN BRAZILIAN WOMEN BY REAL-TIME PCR USING SYBR GREEN I

Abstract

Introduction: The Chlamydia trachomatis represents a major global health challenge as one of the most prevalent sexually transmitted pathogens, with an estimated annual incidence of 100 million cases worldwide. Current diagnostic approaches encompass both conventional and molecular techniques, each demonstrating distinct profiles of analytical sensitivity, specificity, and clinical utility. Objectives: This study aimed to develop and validate a Real-Time PCR-based detection system for C. trachomatis employing SYBR® Green I fluorescence chemistry, offering a balance between diagnostic accuracy and cost-effectiveness. Methods: We analyzed vaginal swab specimens collected from patients attending a women's health clinic in Recife, Brazil. Genomic DNA was isolated using the Wizard® purification system, followed by Real-Time PCR amplification targeting a conserved 150 bp sequence with SYBR® Green I detection. Assay specificity was rigorously evaluated through melt curve analysis (77 ± 0.5°C characteristic peak) and confirmed by agarose gel electrophoresis. Results: Among 98 clinical specimens evaluated, we identified a C. trachomatis infection prevalence of 14.3%. The optimized protocol demonstrated excellent analytical performance with well-defined amplification kinetics and specific melting profiles. Conclusion: Our research assay provides a robust, economical, and technically accessible platform for C. trachomatis screening, particularly suitable for clinical laboratories in resource-conscious settings. The methodology combines the sensitivity of nucleic acid amplification with the cost advantages of intercalating dye chemistry, representing a valuable tool for STI control programs.