Objective To evaluate new test kits and a reverse algorithm for the diagnosis of syphilis at the Queen’s Park Counselling Centre and Clinic (QPCC&C), the national reference laboratory in Trinidad and Tobago, to determine feasibility for implementation. Methods The diagnostic testing for syphilis in Trinidad and Tobago involved the use of the traditional algorithm. The QPCC&C in collaboration with the Centers for Disease Control and Prevention STI & International Laboratory Branches (CDC-STI), Atlanta, developed a reverse diagnostic syphilis algorithm that allows specimens to be screened with a treponemal test (ELISA) and non reactive samples are reported as negative for syphilis. ELISA reactive specimens were reflexively screened with the quantitative VDRL test to detect present or past treated exposure to syphilis. Samples with discordant test results were subsequently screened using the Treponema Pallidum Particle Agglutination (TPPA) as a confirmatory test. To validate the algorithm, 5mls of blood from 40 high risk HIV and STI patients in Trinidad were obtained, the sera separated and split equally to allow for the simultaneous testing and review of results using the reverse testing algorithm at both the QPCC&C and CDC STI laboratories. Ethical approval was granted for this process by the Ministry of Health, Trinidad and Tobago. Results The results obtained showed 100% concordance between laboratories which resulted in the validation of the algorithm. Conclusion The validation of these test kits and reverse algorithm will allow the national STI reference laboratory to increase its throughput and offer an improved service to the population.