A Multiplex PCR assay for a differential diagnostic of rickettsiosis, Lyme disease and scrub typhus

Background & objectives: Coexistence of tick-borne diseases in some regions in Latin America makes the diagnosis difficult due to shared initial signs and symptoms. Rickettsiosis, Lyme disease and recently, scrub typhus are gaining more importance. The objective of this study is to develop a multiplex-PCR assay for a differential diagnosis of rickettsiosis, Lyme disease and scrub typhus. Methods: By using bibliographic and bioinformatic analysis, we identify candidate regions to perform the multiplex- PCR assay for Rickettsia sp., Borrelia burgdorferi and Orientia tsutsugamushi as well as identify optimal melting temperature and sensibility analysis. Results: We identified specific primer pairs for Rickettsia sp, Borrelia burgdorferi and Orientia tsutsugamushi with different PCR fragment length but a common melting temperature, 58°C. Interpretation & conclusion: We successfully developed a Multiplex PCR assay for differential diagnosis of rickettsiosis, Lyme disease and scrub typhus that could be a rapid and easy option in clinical and epidemiological practice.