Analysis of DNA extraction methods for detection of Treponema pallidum: A comparison of three methods

Syphilis is a sexually transmitted disease caused by Treponema pallidum. DNA amplification methods have started to be used to facilitate diagnosis at different stages of the disease. The success of such methodologies depends on obtaining DNA from clinical samples in adequate quantity and quality for molecular reactions. There are many DNA extraction kits, but often the molecular analysis process is unfeasible due to its cost and access to imported products. Thus, this study aimed to analyze three methods of extracting DNA from Treponema pallidum from ulcers of patients investigated for syphilis. The three methods, an in house one (sonication) and two commercial ones (LGC, Brazil) and the PureLink Genomic DNA Mini Kit (Thermo Fisher Scientific, USA) were compared to the sequencing of these samples, which were used as a reference. Each method was evaluated based on the detection of T. pallidum by PCR using the tpp47 gene as a target for amplification, DNA quantification and method execution time. When compared to the sequencing, the sensitivity and agreement of the PureLink, sonication and LGC methods to extracted DNA were 100% (K = 1.0), 96.5% (K = 0.96) and 72.4% (K = 0.694), respectively. Specificity was 100% with the three methods. The sonication method was the closest in concentration of DNA to the PureLink method with a similar degree of purity, besides having the lowest cost-benefit ratio. It can be an interesting option for laboratories that work with reduced costs, since it is much more financially viable.