Development of real-time PCR assays for the specific detection of Francisella tularensis ssp tularensis, holarctica and mediaasiatica

作者：Mitchell Jan L^*; Chatwell Nicola; Christensen Deanna; Diaper Helen; Minogue Timothy D; Parsons Tanya M; Walker Brian; Weller Simon A

来源：Molecular and Cellular Probes, 2010, 24(2): 72-76.

DOI：10.1016/j.mcp.2009.10.004

• Tularaemia
• Real-time PCR
• Diagnosis
• Francisella tularensis

Real-time polymerase chain reaction (PCR) assays were developed to detect Francisella tularensis (Ft), the causative agent of tularaemia in humans. Two real-time PCRs (FTT0376 and FTT0523) were designed in genetic sequences identified by the Insignia genome comparison tool (http://insignia.cbcb.umd.edu/) as being unique to pathogenic subspecies of F tularensis. Both PCRs identified all pathogenic E tularensis subspecies but did not cross react with avirulent Francisella philomiragia or F tularensis ssp. novicida or other environmental bacteria. Limits of detection from DNA purified from pure culture (FTT0376 similar to 80 Ft genome equivalents (GEs) per PCR; FTT0523 similar to 20 Ft GEs per PCR;) and DNA purified from spiked blood samples (4 x 10(4) to 4 x 10(3) cfu ml(-1), both assays) were determined.

• 出版日期2010-4