Comparative evaluation of Panther Fusion and real-time PCR for detection of Burkholderia pseudomallei in spiked human blood

Introduction. Melioidosis is an infection that most commonly presents with bacteraemia. Culture-based laboratory methods can result in a significant delay to organism identification. Molecular diagnostic techniques have a high sensitivity and rapid time to diagnosis. A decreased time to diagnosis is likely to improve patient outcomes. Aim. To compare the Panther Fusion automated molecular instrument to an in-house method for the detection of Burkholderia pseudomallei directly from spiked human whole-blood samples. Results. The in-house method detected 11/12 (92 %) samples with a B. pseudomallei concentration of 2.5–4.5×102 c.f.u. ml−1. The Panther was less reliable, detecting only 8/14 (75 %) samples with a similar bacterial concentration. The Panther was able to detect 12/12 (100 %) spiked blood culture-positive samples. Conclusion. The direct detection of B. pseudomallei from patient blood on presentation to a healthcare facility will significantly decrease time to diagnosis. We describe an in-house real-time PCR method with the lowest reported limit of detection to date. Due to lower sensitivity, the Panther Fusion would be best used as a diagnostic method directly from a positive blood culture.

[1]  M. Chatfield,et al.  The epidemiology of melioidosis in Townsville, Australia. , 2021, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[2]  R. Price,et al.  The Darwin Prospective Melioidosis Study: a 30-year prospective, observational investigation. , 2021, The Lancet. Infectious diseases.

[3]  D. Limmathurotsakul,et al.  A multi-country study using MALDI-TOF mass spectrometry for rapid identification of Burkholderia pseudomallei , 2021, BMC microbiology.

[4]  P. Harris,et al.  Diagnosis of melioidosis: the role of molecular techniques. , 2021, Future microbiology.

[5]  S. Ombelet,et al.  Nonautomated Blood Cultures in a Low-Resource Setting: Optimizing the Timing of Blind Subculture , 2020, The American journal of tropical medicine and hygiene.

[6]  K. Stellrecht,et al.  The Panther Fusion System with Open Access Functionality for Laboratory-Developed Tests for Influenza A Virus Subtyping , 2020, Journal of Clinical Microbiology.

[7]  R. Norton,et al.  Human Melioidosis , 2020, Clinical Microbiology Reviews.

[8]  N. Day,et al.  Clinical Epidemiology of 7126 Melioidosis Patients in Thailand and the Implications for a National Notifiable Diseases Surveillance System , 2019, Open forum infectious diseases.

[9]  E. Bertherat,et al.  Global burden of melioidosis in 2015: a systematic review and data synthesis. , 2019, The Lancet. Infectious diseases.

[10]  R. Norton,et al.  Identification of Burkholderia pseudomallei by Use of the Vitek Mass Spectrometer , 2019, Journal of Clinical Microbiology.

[11]  Hyunwoong Park,et al.  Comparison of Clinical Performance Between BacT/Alert Virtuo and BacT/Alert 3D Blood Culture Systems , 2019, Annals of laboratory medicine.

[12]  Sarah K. Parker,et al.  Implementation of Rapid Molecular Infectious Disease Diagnostics: the Role of Diagnostic and Antimicrobial Stewardship , 2016, Journal of Clinical Microbiology.

[13]  S. Hay,et al.  Predicted global distribution of Burkholderia pseudomallei and burden of melioidosis , 2016, Nature Microbiology.

[14]  A. Hoffmaster,et al.  Comparison of DNA Extraction Kits for Detection of Burkholderia pseudomallei in Spiked Human Whole Blood Using Real-Time PCR , 2013, PloS one.

[15]  Talima Pearson,et al.  Comparison of TaqMan PCR Assays for Detection of the Melioidosis Agent Burkholderia pseudomallei in Clinical Specimens , 2012, Journal of Clinical Microbiology.

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

[17]  Eileen M. Burd,et al.  Validation of Laboratory-Developed Molecular Assays for Infectious Diseases , 2010, Clinical Microbiology Reviews.

[18]  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.

[19]  N. Day,et al.  Quantitation of B. Pseudomallei in clinical samples. , 2007, The American journal of tropical medicine and hygiene.

[20]  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.

[21]  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.

[22]  C. Hart,et al.  Evidence for the presence in Burkholderia pseudomallei of a type III secretion system-associated gene cluster. , 1999, Journal of medical microbiology.

[23]  T. Dharakul,et al.  Speed of detection of Burkholderia pseudomallei in blood cultures and its correlation with the clinical outcome. , 1997, The American journal of tropical medicine and hygiene.

[24]  A. E. Lew,et al.  Detection of Pseudomonas pseudomallei by PCR and hybridization , 1994, Journal of clinical microbiology.