Recombinase Polymerase Amplification Assay for Rapid Detection of Francisella tularensis

ABSTRACT Several real-time PCR approaches to develop field detection for Francisella tularensis, the infectious agent causing tularemia, have been explored. We report the development of a novel qualitative real-time isothermal recombinase polymerase amplification (RPA) assay for use on a small ESEQuant Tube Scanner device. The analytical sensitivity and specificity were tested using a plasmid standard and DNA extracts from infected rabbit tissues. The assay showed a performance comparable to real-time PCR but reduced the assay time to 10 min. The rapid RPA method has great application potential for field use or point-of-care diagnostics.

[1]  Yi Zhang,et al.  Advances in microfluidic PCR for point-of-care infectious disease diagnostics. , 2011, Biotechnology advances.

[2]  Felicie F. Andersen,et al.  Strategies for highly sensitive biomarker detection by Rolling Circle Amplification of signals from nucleic acid composed sensors. , 2011, Integrative biology : quantitative biosciences from nano to macro.

[3]  Peter J. Asiello,et al.  Miniaturized isothermal nucleic acid amplification, a review. , 2011, Lab on a chip.

[4]  Christopher J Easley,et al.  Isothermal DNA amplification in bioanalysis: strategies and applications. , 2011, Bioanalysis.

[5]  P. Kämpfer,et al.  Description of Francisella hispaniensis sp. nov., isolated from human blood, reclassification of Francisella novicida (Larson et al. 1955) Olsufiev et al. 1959 as Francisella tularensis subsp. novicida comb. nov. and emended description of the genus Francisella. , 2010, International journal of systematic and evolutionary microbiology.

[6]  Roland Zengerle,et al.  Microfluidic lab-on-a-foil for nucleic acid analysis based on isothermal recombinase polymerase amplification (RPA). , 2010, Lab on a chip.

[7]  Raymond K. Auerbach,et al.  Francisella tularensis subsp. novicida isolated from a human in Arizona , 2009, BMC Research Notes.

[8]  H. Kong,et al.  Development of isothermal TaqMan assays for detection of biothreat organisms. , 2008, BioTechniques.

[9]  P. Mead,et al.  Francisella tularensis: an arthropod-borne pathogen , 2008, Veterinary research.

[10]  P. Gill,et al.  Nucleic Acid Isothermal Amplification Technologies—A Review , 2008, Nucleosides, nucleotides & nucleic acids.

[11]  Olaf Piepenburg,et al.  DNA Detection Using Recombination Proteins , 2006, PLoS biology.

[12]  P. Schuff-Werner,et al.  Diagnostic procedures in tularaemia with special focus on molecular and immunological techniques. , 2005, Journal of veterinary medicine. B, Infectious diseases and veterinary public health.

[13]  Yan Xu,et al.  Helicase‐dependent isothermal DNA amplification , 2004, EMBO reports.

[14]  L. Lemming,et al.  Problems in Identification of Francisella philomiragia Associated with Fatal Bacteremia in a Patient with Chronic Granulomatous Disease , 2004, Journal of Clinical Microbiology.

[15]  Jeannine M. Petersen,et al.  Development of a Multitarget Real-Time TaqMan PCR Assay for Enhanced Detection of Francisella tularensis in Complex Specimens , 2003, Journal of Clinical Microbiology.

[16]  David M. Hartley,et al.  Syndromic Surveillance and Bioterrorism-related Epidemics , 2003, Emerging infectious diseases.

[17]  F. Hufert,et al.  Rapid Detection of Human Pathogenic Orthobunyaviruses , 2003, Journal of Clinical Microbiology.

[18]  Frank T. Hufert,et al.  Rapid Detection of Herpes Simplex Virus and Varicella-Zoster Virus Infections by Real-Time PCR , 2003, Journal of Clinical Microbiology.

[19]  Peter A. Emanuel,et al.  Detection of Francisella tularensis within Infected Mouse Tissues by Using a Hand-Held PCR Thermocycler , 2003, Journal of Clinical Microbiology.

[20]  Mats Nilsson,et al.  Real-time monitoring of rolling-circle amplification using a modified molecular beacon design. , 2002, Nucleic acids research.

[21]  J. Power,et al.  Strategies for signal amplification in nucleic acid detection , 2001, Molecular biotechnology.

[22]  Philip K. Russell,et al.  Tularemia as a biological weapon: medical and public health management. , 2001, JAMA.

[23]  C. Goldsmith,et al.  Replicate PCR Testing and Probit Analysis for Detection and Quantitation of Chlamydia pneumoniae in Clinical Specimens , 2001, Journal of Clinical Microbiology.

[24]  F. Kramer,et al.  Molecular beacon probes combined with amplification by NASBA enable homogeneous, real-time detection of RNA. , 1998, Nucleic acids research.

[25]  F. Polack,et al.  Recurrent Francisella philomiragia sepsis in chronic granulomatous disease. , 1998, The Pediatric infectious disease journal.

[26]  S. Sicherer,et al.  Francisella philomiragia sepsis in chronic granulomatous disease. , 1997, The Pediatric infectious disease journal.

[27]  C. Broome,et al.  Infection caused by Francisella philomiragia (formerly Yersinia philomiragia). A newly recognized human pathogen. , 1989, Annals of internal medicine.

[28]  L. Nuotio,et al.  Rapid field detection assays for Bacillus anthracis, Brucella spp., Francisella tularensis and Yersinia pestis. , 2011, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[29]  D. Norwood,et al.  Detection of biological threat agents by real-time PCR: comparison of assay performance on the R.A.P.I.D., the LightCycler, and the Smart Cycler platforms. , 2006, Clinical chemistry.

[30]  C. Batt,et al.  A microchip-based DNA purification and real-time PCR biosensor for bacterial detection , 2004, Proceedings of IEEE Sensors, 2004..