Tick-borne rickettsioses are worldwide infectious diseases that are considered emerging and re-emerging. Until recently the only tick-borne rickettsiosis present in Latin America was Rickettsia rickettsii infection, but to date, with the incorporation of new tools as PCR and sequencing and the quick cellular close tube cultures (Shell-vial), new species has been involved as human pathogens. In these guidelines, we offer an update of the microbiological assays for diagnosing rickettsioses. Besides we have included a section in which the most important hard ticks involved in human rickettsioses in Latinoamerica are detailed.