Syphilis, caused by the bacterium Treponema pallidum, is on the rise in the United States particularly among men who have sex with men. The disease is complex with varied clinical manifestations and challenges remain in the laboratory diagnostic setting because T. pallidum is noncultivable and no single test can accurately diagnose all stages of the disease. There are missed opportunities for the use of direct detection tests in primary and secondary syphilis. The increasing use of different reverse sequence algorithms for serology testing without validation in populations with varying risks for syphilis makes the interpretation of test results difficult; this has led to concerns about diagnostic errors or overtreatment. On the other hand, the traditional algorithm may miss some early primary syphilis cases, which is of concern in high-risk populations. The potential utility of rapid syphilis serology tests in different settings or populations remains to be determined. The implementation of better tests and appropriate testing algorithms together with laboratory guidelines for test use in general will lead to better diagnostic options for syphilis.