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A HIGHLY SENSITIVE AND SPECIFIC REAL-TIME PCR ASSAY FOR THE DETECTION OF SPOTTED FEVER AND TYPHUS GROUP RICKETTSIAE

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.

Received March 14, 2005. Accepted for publication August 19, 2005.

Acknowledgments: We would like to thank The Geelong Region Medical Research Foundation for financial assistance with this project and Lin Dillon from Westmead Hospital, Sydney, for supplying the Bartonella strains.

^* Address correspondence to Nathan Unsworth, The Australian Rickettsial Reference Laboratory, The Geelong Hospital, P.O. Box 281, Geelong, Victoria, Australia 3220. E-mail: nathanu{at}barwonhealth.org.au

Authors’ addresses: John Stenos, Stephen R. Graves, and Nathan B. Unsworth, The Australian Rickettsial Reference Laboratory, The Geelong Hospital, P.O. Box 281, Geelong, Victoria, Australia 3220, Telephone: 61 3 5226 7521, Fax: 61 3 5260 3183.

Reprint requests: Nathan Unsworth, The Australian Rickettsial Reference Laboratory, The Geelong Hospital, P.O. Box 281, Geelong, Victoria, Australia 3220, Telephone: 61-3-5226-7521, Fax: 61-3-5260-3183, E-mail: nathanu{at}barwonhealth.org.au.

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