Real-time multiplex PCR assay for detection and differentiation of rickettsiae and orientiae》.pdf

Transactions of the Royal Society of Tropical Medicine and Hygiene (2008) 102, 186—193 a v a i la b le at ww w.scie ncedirect. com j o u rn al h omepage: www.elsevi /journals/tr st Real-time multiplex PCR assay for detection and differentiation of rickettsiae and orientiae Daniel H. Parisa,b,∗, Stuart D. Blacksella,b, John Stenosc, Stephen R. Graves c, Nathan B. Unsworthc,d, Rattanaphone Phetsouvanhe, Paul N. Newton b,e, Nicholas P.J. Daya,b a Mahidol—Oxford Tropical Medicine Programme, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand b Centre for Tropical Medicine, Nuffield Department of Clinical Medicine, Churchill Hospital, University of Oxford, Oxford, UK c The Australian Rickettsial Reference Laboratory, Department of Clinical and Biomedical Sciences, The University of Melbourne, The Geelong Hospital, Victoria, Australia d The Department of Microbial and Molecular Pathogenesis, The College of Medicine, Texas AM University, Texas, USA e Wellcome Trust—Mahosot Hospital—Oxford Tropical Medicine Research Collaboration, Mahosot Hospital, Vientiane, Lao PDR Received 24 April 2007; received in revised form 1 November 2007; accepted 1 November 2007 KEYWORDS Summary The high incidence of rickettsial diseases in Southeast Asia necessitates rapid Orientia and accurate diagnostic tools for a broad range of rickettsial agents, including Orientia tsut- tsutsugamushi; sugamushi (scrub typhus) and Rickettsia typhi (murine typhus), but also spotted fever group Rickettsia typhi; infections, which are increasingly reported. We present an SYBR-Green-based, real-time multi- plex PCR assay for rapid identification and differentiation of scrub typhus group, typhus group Diagnosis;