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GNR Traffic Light™ PNA FISH ®

GNR Traffic Light™ PNA FISH ®. Rapid Identification of E. coli , K. pneumoniae and P. aeruginosa from Positive Blood Cultures. GNR Traffic Light ™ PNA FISH ® . 90 min. identification and differentiation of E. coli , K. pneumoniae and P. aeruginosa from GNR-positive blood cultures

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GNR Traffic Light™ PNA FISH ®

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  1. GNR Traffic Light™ PNA FISH® Rapid Identification of E. coli, K. pneumoniae and P. aeruginosa from Positive Blood Cultures PN1874A

  2. GNR Traffic Light™ PNA FISH® • 90 min. identification and differentiation of E. coli, K. pneumoniae and P. aeruginosa from GNR-positive blood cultures • Help optimize antibiotic therapy (Pseudomonal vs. non-Pseudomonal) 1-2 days earlier for Gram-Negative bacteremia • Help improve clinical outcomes while controlling antibiotic use Gram Stain PNA FISH® in 90 Minutes GNR E. coli (35%) K. pneumoniae (20%) P. aeruginosa (15%) PN1874A

  3. Performance Data Includes 4 samples spiked with P. aeruginosa clinical isolates. One false negative for K. pneumoniae in a mixed culture of E. coli and K. pneumoniae. Both false negatives were in mixed cultures of P. aeruginosa and K. pneumoniae. One false green positive E. cloacae (negative upon re-test) and one false red positive A. baumannii. One false red positive Acinetobacter radioresistensin a mixed culture with E. faecalis Includes 28 samples spiked with P. aeruginosa isolates. Please see Package Insert for complete performance data and limitations PN1874A

  4. Bloodstream Infections (BSIs) • Bacteremia and Candidemia are serious infections1 • Mortality: 14% (community) – 34% (nosocomial). • >50% of bloodstream infections are hospital-acquired (nosocomial). • Condition with highest increase in incidence from 2001 to 20072 • 675,000 annual cases in US2 • Increase of 97% from 2001 to 2007. • Costs = $12.3 billion ($18.2K per patient)2 • Increase of 174% from 2001 to 2007. $12.3B 675,000 342,500 $4.5B • Diekema et al. J Clin Microbiol. 2003 Aug;41(8):3655-60. • Cost of Hospital Treatment for Blood Infection Surges, Especially for Uninsured Patients. AHRQ News and Numbers, June 9, 2010. Agency for Healthcare Research and Quality, Rockville, MD. http://www.ahrq.gov/news/nn/nn060910.htm PN1874A

  5. Early, appropriate therapy is key to survival n = 2,731 Kumar et al. Duration of hypotension before initiation of effective antimicrobial therapy is the critical determinant of survival in human septic shock. Crit Care Med. 2006 Jun;34(6):1589-96. PN1874A

  6. Inappropriate therapy is common Major Pathogens with Highest Rates of Inappropriate Therapy (Severe Sepsis) Accounted for 61% of episodes Occurrence of inappropriate therapy n = 1,901 n = 1,901 Golan et al. Candidal vs. Bacterial Severe Sepsis in the ICU: The Number-Needed-to-Treat (NNT). Poster # 631. IDSA 2010. Vancouver, BC, Canada. PN1874A

  7. Systemic infections – therapy balancing act • Ensure early, appropriate and effective antibiotic therapy for patients with critical infections. • Avoid overexposure and unnecessary use of broad-spectrum antibiotics in lieu of rising resistance and lack of new antibiotics. Adapted from Centers from Diseases Control and Prevention data Spellberg et al. The epidemic of antibiotic-resistant infections: a call to action for the medical community from the Infectious Diseases Society of America. Clin Infect Dis. 2008 Jan 15;46(2):155-64. PN1874A

  8. Time is critical for positive blood cultures You need results… Now “If you were septic, would you wait 48-72 hours to know if you are receiving appropriate or unnecessary antibiotic therapy?” Dr. Graeme Forrest. Infectious Diseases. University of Maryland Medical CenterICAAC 2006, San Francisco, CA PN1874A

  9. PNA FISH: Pathogen ID in 90 minutes PositiveBlood Culture Conventional Species ID Gram Stain Blood Draw Conventional Dx Empiric Rx Broad-spectrum Rx Targeted Rx ...Rapid and accurate results on day 1: • Supports decisions for appropriate and targeted therapy 1-3 days earlier than conventional methods PositiveBlood Culture Blood Draw Gram Stain Species ID PNA FISH® Empiric Rx Targeted Rx PN1874A

  10. PNA probes target ribosomal RNA inside cells Peptide Nucleic Acid Fluorescence In Situ Hybridization PN1874A

  11. PNA probes – Peptide Nucleic Acid • Fast, specific and tight binding to RNA targets. • No electrostatic repulsion with charged RNA backbone. • rRNA sequences are highly conserved and well defined • Ideal targets for phylogenetic analysis (i.e. species ID) • rRNA content high in a growing bacteria. • “Naturally amplified” in viable cells • No need for any further amplification. PN1874A

  12. Whole Cell Analysis (WCA) • Cells kept intact • PNA probes hybridize to rRNA inside bacteria enabling whole cell analysis • Retains morphology – similar to Gram stain • No need to lyse cells, allowing for simple sample preparation • Cells can be counted • Quantification: utility in future application such as testing of respiratory and urine samples • Multiplexing by color and shape PN1874A

  13. Gram stain selects PNA FISH tests PositiveBlood Culture(8-12% Positive) Gram Stain PNA FISH® in 90 Minutes ~55% CNS GPCC S. aureus CNS Mixed Positive ~15% GPCPC E. faecalis E. faecium & OE Mixed Positive ~20% E. coli K. pneumonia P. aeruginosa GNR ~5% C. albicans and/orC. parapsilosis C. tropicalis C. glabrata and/orC. krusei Yeast PN1874A

  14. PNA FISH® – Rapid, Simple & Easy Protocol Prepare Smear Hybridize Wash Examine 5-20 min.* 30 min. 30 min. View Results • Add drop of Fixation Solution • Add drop from BC+ • Fix bacteria/yeast onto slide • Heat • Methanol, or • Flame fixation • Add PNA Probe • Probe enters cells and binds to target rRNA sequence, if present • Immerse slide in Wash Solution • Unbound and excess PNA Probe removed from cells and slide • Fluorescence microscopy using 60x or 100x oil objective • Target bacteria/yeast fluoresce Turn Around Time ≤ 90 min.Hands-On Time = 10 min. Please see package insert for complete instructions for use *Dependent on fixation method PN1874A

  15. Limited Capital Start-Up PNA FISH Starter System(Equipment Required for Performing PNA FISH) Fluorescence Microscope(60x or 100x Oil Objective) PN1874A

  16. Bloodstream Infections: Gram Stain Dilemmas Yeast: Fluconazole vs. Echinocandin? GPCC: True S. aureus Infection vs. CNS Contamination? GNR: P. aeruginosa Rx vs. non-P. aeruginosa Rx? GPCPC: Ampicillin and Vancomycin Resistant E. faecium? Karlowsky et al. Prevalence and antimicrobial susceptibilities of bacteria isolated from blood cultures of hospitalized patients in the United States in 2002. Ann Clin Microbiol Antimicrob. 2004 May 10;3:7. PN1874A

  17. Gram-Negatives in Blood Cultures • Gram stain: Gram-Negative Rods • Cannot distinguish E. coli, K. pneumoniae, or P. aeruginosa from other GNRs. • Dilemma: Is the Gram-negative rods in the blood culture Multi-Drug Resistant or not? • When to cover for P. aeruginosa? • When to cover for KPC? • When to de-escalate from anti-pseudomonal therapy? • Conventional culture identification takes 1-2 days. • Clinicians can’t wait. They need to treat. Pseudomonal vs. Non-Pseudomonal Rx? PN1874A

  18. Consequences • Delayed administration of appropriate therapy for patients with P. aeruginosa bacteremia • Higher mortality • Significant extra hospital cost • Broader-than-necessary antibiotic therapy for patients with bloodstream infections caused by other GNRs • Selection for MDR pathogens • Risk of toxicity complications 74% decrease Micek ST et al. Pseudomonas aeruginosa bloodstream infection: importance of appropriate initial antimicrobial treatment. Antimicrob Agents Chemother. 2005 Apr;49(4):1306-11. Kumar et al. Initiation of Inappropriate Antimicrobial Therapy Results in a 5-Fold Reduction of Survival in Human Septic Shock. Chest. 2009 Nov;136(5):1237-48. Epub 2009 Aug 20. PN1874A

  19. Early, appropriate therapy for P. aeruginosa bacteremia is critical for survival 72% increase Micek ST et al. Pseudomonas aeruginosa bloodstream infection: importance of appropriate initial antimicrobial treatment. Antimicrob Agents Chemother. 2005 Apr;49(4):1306-11. Kang et al. Pseudomonas aeruginosa Bacteremia: Risk Factors for Mortality and Influence of Delayed Receipt of Effective Antimicrobial Therapy on Clinical Outcome. Clin Infect Dis. 2003; 37: 745-51 PN1874A

  20. GNRs in Blood Cultures: The Challenge The challenge for clinicians is to decide when to initiate or discontinue anti-pseudomonal therapy Karlowsky et al. Prevalence and antimicrobial susceptibilities of bacteria isolated from blood cultures of hospitalized patients in the United States in 2002. Ann Clin Microbiol Antimicrob. 2004 May 10;3:7. PN1874A

  21. Conclusion: GNR Traffic Light™ PNA FISH® • Identifies and differentiates between E. coli, K. pneumoniae and P. aeruginosa from GNR-positive blood cultures. • Results 1-2 days earlier than conventional methods. • Aid in optimizing antibiotic therapy earlier for Gram-Negative bacteremia. • Support clinician’s decision on when to initiate or discontinue anti-pseudomonal therapy to improve clinical outcomes. PN1874A

  22. GNR Traffic Light™ PNA FISH® PN1874A

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