Comparison of TaqMan PCR Assays for Detection of the Melioidosis Agent Burkholderia pseudomallei in Clinical Specimens

ABSTRACT Melioidosis is an emerging infectious disease caused by the soil bacterium Burkholderia pseudomallei. In diagnostic and forensic settings, molecular detection assays need not only high sensitivity with low limits of detection but also high specificity. In a direct comparison of published and newly developed TaqMan PCR assays, we found the TTS1-orf2 assay to be superior in detecting B. pseudomallei directly from clinical specimens. The YLF/BTFC multiplex assay (targeting the Yersinia-like fimbrial/Burkholderia thailandensis-like flagellum and chemotaxis region) also showed high diagnostic sensitivity and provides additional information on possible geographic origin.

[1]  D. Armbruster,et al.  Limit of blank, limit of detection and limit of quantitation. , 2008, The Clinical biochemist. Reviews.

[2]  D. Engelthaler,et al.  BurkDiff: A Real-Time PCR Allelic Discrimination Assay for Burkholderia Pseudomallei and B. mallei , 2010, PloS one.

[3]  J. Schrenzel,et al.  Fatal acute melioidosis in a tourist returning from Martinique Island, November 2010. , 2011, Euro surveillance : bulletin Europeen sur les maladies transmissibles = European communicable disease bulletin.

[4]  A. Cheng,et al.  The global distribution of Burkholderia pseudomallei and melioidosis: an update. , 2008, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[5]  Nicholas J Loman,et al.  High-throughput sequencing and clinical microbiology: progress, opportunities and challenges. , 2010, Current opinion in microbiology.

[6]  S. Panyim,et al.  A simple method to detect and differentiate Burkholderia pseudomallei and Burkholderia thailandensis using specific flagellin gene primers. , 2002, Molecular and cellular probes.

[7]  V. Beneš,et al.  The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. , 2009, Clinical chemistry.

[8]  J. Schrenzel,et al.  Application and use of various mass spectrometry methods in clinical microbiology. , 2010, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[9]  D. Engelthaler,et al.  Towards a rapid molecular diagnostic for melioidosis: Comparison of DNA extraction methods from clinical specimens. , 2012, Journal of microbiological methods.

[10]  T. Panichakul,et al.  Monoclonal antibody-based rapid identification of Burkholderia pseudomallei in blood culture fluid from patients with community-acquired septicaemia. , 2000, Journal of medical microbiology.

[11]  W. Chantratita,et al.  Prospective clinical evaluation of the accuracy of 16S rRNA real-time PCR assay for the diagnosis of melioidosis. , 2007, The American journal of tropical medicine and hygiene.

[12]  T. Inglis,et al.  PCR-based identification of Burkholderia pseudomallei. , 2006, Revista do Instituto de Medicina Tropical de Sao Paulo.

[13]  R. Dargis,et al.  Detection of Burkholderia pseudomallei by SYBR Green Real Time PCR , 2009 .

[14]  D. Vidal,et al.  Identification and Discrimination of Burkholderia pseudomallei, B. mallei, and B. thailandensis by Real-Time PCR Targeting Type III Secretion System Genes , 2004, Journal of Clinical Microbiology.

[15]  Ryan T Novak,et al.  Clinical Evaluation of a Type III Secretion System Real-Time PCR Assay for Diagnosing Melioidosis , 2006, Journal of Clinical Microbiology.

[16]  Ryan T Novak,et al.  Development and Evaluation of a Real-Time PCR Assay Targeting the Type III Secretion System of Burkholderia pseudomallei , 2006, Journal of Clinical Microbiology.

[17]  D. Dance,et al.  Melioidosis as an emerging global problem. , 2000, Acta tropica.

[18]  A. Hensel,et al.  Development and Clinical Evaluation of A PCR Assay Targeting the Metalloprotease Gene (mprA) of B. pseudomallei , 2007, Zoonoses and public health.

[19]  A. Cheng,et al.  Evaluation of Immunoglobulin M (IgM) and IgG Rapid Cassette Test Kits for Diagnosis of Melioidosis in an Area of Endemicity , 2004, Journal of Clinical Microbiology.

[20]  Raymond K. Auerbach,et al.  A Horizontal Gene Transfer Event Defines Two Distinct Groups within Burkholderia pseudomallei That Have Dissimilar Geographic Distributions , 2007, Journal of bacteriology.

[21]  P. Newton,et al.  Accuracy of Burkholderia pseudomallei Identification Using the API 20NE System and a Latex Agglutination Test , 2007, Journal of Clinical Microbiology.

[22]  P. Woo,et al.  Single gene target bacterial identification. groEL gene sequencing for discriminating clinical isolates of Burkholderia pseudomallei and Burkholderia thailandensis. , 2002, Diagnostic microbiology and infectious disease.

[23]  A. Cheng,et al.  Melioidosis: Epidemiology, Pathophysiology, and Management , 2005, Clinical Microbiology Reviews.

[24]  J. Vadivelu,et al.  Development and evaluation of polymerase chain reaction assay to detect Burkholderia genus and to differentiate the species in clinical specimens. , 2010, FEMS microbiology letters.

[25]  DNA microarray-based detection and identification of Burkholderia mallei, Burkholderia pseudomallei and Burkholderia spp. , 2009, Molecular and cellular probes.

[26]  A. Cheng,et al.  Rapid Immunofluorescence Microscopy for Diagnosis of Melioidosis , 2005, Clinical Diagnostic Laboratory Immunology.

[27]  Tanja Popovic,et al.  Use of 16S rRNA Gene Sequencing for Rapid Identification and Differentiation of Burkholderia pseudomallei and B. mallei , 2003, Journal of Clinical Microbiology.

[28]  B. Currie Melioidosis: an important cause of pneumonia in residents of and travellers returned from endemic regions , 2003, European Respiratory Journal.

[29]  N. Day,et al.  Loop-Mediated Isothermal Amplification Method Targeting the TTS1 Gene Cluster for Detection of Burkholderia pseudomallei and Diagnosis of Melioidosis , 2007, Journal of Clinical Microbiology.

[30]  A. Cheng,et al.  Indirect hemagglutination assay in patients with melioidosis in northern Australia. , 2006, The American journal of tropical medicine and hygiene.

[31]  R. Norton,et al.  Comparison of Automated and Nonautomated Systems for Identification of Burkholderia pseudomallei , 2002, Journal of Clinical Microbiology.

[32]  Claudio J. Verzilli,et al.  Defining the True Sensitivity of Culture for the Diagnosis of Melioidosis Using Bayesian Latent Class Models , 2010, PloS one.

[33]  T. Pitt,et al.  Rapid presumptive identification of Burkholderia pseudomallei with real-time PCR assays using fluorescent hybridization probes. , 2005, Molecular and cellular probes.

[34]  E. Ooi,et al.  Development of Real-Time PCR Assays and Evaluation of Their Potential Use for Rapid Detection of Burkholderia pseudomallei in Clinical Blood Specimens , 2007, Journal of Clinical Microbiology.