title: A highly sensitive and specific real-time PCR assay for the detection of spotted fever and typhus group Rickettsiae.
abstract: A highly specific real-time polymerase chain reaction (PCR) assay was developed to detect spotted fever and typhus group rickettsiae using the citrate synthase gene as the target. The assay amplified rickettsial members of the spotted fever and typhus group including Rickettsia akari, R. australis, R. conorii, R. honei, "R. marmionii," R. sibirica, R. rickettsii, R. typhi, and R. prowazekii. The ancestral group rickettsia, R. bellii, did not produce a positive reaction, nor did other members of the order Rickettsiales or any non-rickettsial bacteria. The assay had a sensitivity of one target copy number per reaction as determined by serial dilutions of a plasmid containing a spotted fever group target sequence. This quantitative assay is useful for the enumeration of rickettsiae in clinical specimens and the diagnosis of rickettsial illnesses, when rickettsial numbers are very low.
authors.affiliation: The Australian Rickettsial Reference Laboratory, Department of Clinical and Biomedical Sciences, The University of Melbourne, The Geelong Hospital, Victoria, Australia.
authors.forename: John
authors.initials: J
authors.lastname: Stenos
authors.forename: Stephen R
authors.initials: SR
authors.lastname: Graves
authors.forename: Nathan B
authors.initials: NB
authors.lastname: Unsworth
mesh_terms: D002950:Citrate (si)-Synthase / Q000235:genetics; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis; D006801:Humans; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D011933:Reagent Kits, Diagnostic; D012281:Rickettsia / Q000145:classification* / Q000235:genetics; D012282:Rickettsia Infections / Q000175:diagnosis* / Q000382:microbiology / Q000635:transmission; D012373:Rocky Mountain Spotted Fever / Q000175:diagnosis / Q000382:microbiology / Q000635:transmission; D012680:Sensitivity and Specificity; D014739:Victoria / Q000453:epidemiology
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D011933:Reagent Kits, Diagnostic; D002950:Citrate (si)-Synthase