New Real-Time PCRs to Differentiate Rickettsia spp. and Rickettsia conorii.

Rickettsia species are an important cause of emerging infectious diseases in people and animals, and rickettsiosis is one of the oldest known vector-borne diseases. Laboratory diagnosis of Rickettsia is complex and time-consuming. This study was aimed at developing two quantitative real-time PCRs targeting ompB and ompA genes for the detection, respectively, of Rickettsia spp. and R. conorii DNA. Primers were designed following an analysis of Rickettsia gene sequences. The assays were optimized using SYBR Green and TaqMan methods and tested for sensitivity and specificity. This study allowed the development of powerful diagnostic methods, able to detect and quantify Rickettsia spp. DNA and differentiate R. conorii species.

A retrospective study of the characterization of Rickettsia species in ticks collected from humans.

Rickettsiae (family Rickettsiaceae, order Rickettsiales) are obligate intracellular bacteria transmitted by arthropod vectors. Several Rickettsia species causing vector-borne rickettsioses belong to the spotted fever group (SFG). Traditionally, Rickettsia conorii has been considered as the main etiologic agent of Mediterranean spotted fever. However, the molecular characterization of rickettsiae allowed identifying other species involved in spotted fever in the Mediterranean region. In this study, 42 ticks collected from humans were subjected to morphological identification and molecular characterization of Rickettsia species potentially involved in human rickettsiosis in Sicily. Fourteen ticks positive to at least two Rickettsia spp. molecular markers were used in the study. Identified Rickettsia spp. included R. conorii, found in Rhipicephalus sanguineus sensu lato and Rhipicephalus turanicus, Rickettsia aeschlimannii found in Hyalomma marginatum, Hyalomma lusitanicum, Dermacentor marginatus and Ixodes ricinus, Rickettsia massiliae found in R. turanicus and R. sanguineus s.l., and Rickettsia slovaca found in D. marginatus and R. sanguineus s.l. Our results showed a great variety of zoonotic Rickettsia spp. in ticks collected from humans in Sicily. The Rickettsia spp. reported in this study were identified in previously recognized or new potential tick vectors in Europe, highlighting the risk of infection by different Rickettsia spp. for humans bitten by ticks in Sicily.