scholarly journals Duplex DNA-Invading γ-Modified Peptide Nucleic Acids Enable Rapid Identification of Bloodstream Infections in Whole Blood

mBio ◽  
2016 ◽  
Vol 7 (2) ◽  
Author(s):  
Jörk Nölling ◽  
Srinivas Rapireddy ◽  
Joel I. Amburg ◽  
Elizabeth M. Crawford ◽  
Ranjit A. Prakash ◽  
...  
Keyword(s):  
Blood Culture ◽  
Nucleic Acids ◽  
Fungal Pathogens ◽  
Peptide Nucleic Acids ◽  
Unmet Need ◽  
Bloodstream Infections ◽  
Rapid Identification ◽  
Duplex Dna ◽  
Diagnostic Approaches ◽  
Antimicrobial Intervention

ABSTRACT Bloodstream infections are a leading cause of morbidity and mortality. Early and targeted antimicrobial intervention is lifesaving, yet current diagnostic approaches fail to provide actionable information within a clinically viable time frame due to their reliance on blood culturing. Here, we present a novel pathogen identification (PID) platform that features the use of duplex DNA-invading γ-modified peptide nucleic acids (γPNAs) for the rapid identification of bacterial and fungal pathogens directly from blood, without culturing. The PID platform provides species-level information in under 2.5 hours while reaching single-CFU-per-milliliter sensitivity across the entire 21-pathogen panel. The clinical utility of the PID platform was demonstrated through assessment of 61 clinical specimens, which showed >95% sensitivity and >90% overall correlation to blood culture findings. This rapid γPNA-based platform promises to improve patient care by enabling the administration of a targeted first-line antimicrobial intervention. IMPORTANCE Bloodstream infections continue to be a major cause of death for hospitalized patients, despite significant improvements in both the availability of treatment options as well their application. Since early and targeted antimicrobial intervention is one of the prime determinants of patient outcome, the rapid identification of the pathogen can be lifesaving. Unfortunately, current diagnostic approaches for identifying these infections all rely on time-consuming blood culture, which precludes immediate intervention with a targeted antimicrobial. To address this, we have developed and characterized a new and comprehensive methodology, from patient specimen to result, for the rapid identification of both bacterial and fungal pathogens without the need for culturing. We anticipate broad interest in our work, given the novelty of our technical approach combined with an immense unmet need.

scholarly journals Evaluation of the ePlex Blood Culture Identification Panels for Detection of Pathogens in Bloodstream Infections

2018 ◽  
Vol 57 (2) ◽  
Author(s):  
Te-Din Huang ◽  
Ekaterina Melnik ◽  
Pierre Bogaerts ◽  
Stephanie Evrard ◽  
Youri Glupczynski
Keyword(s):  
Blood Culture ◽  
Positive Blood Culture ◽  
Fungal Pathogens ◽  
Bacterial Resistance ◽  
Bloodstream Infections ◽  
Rapid Identification ◽  
Added Value ◽  
Gram Positive ◽  
Gram Negative ◽  
Culture Identification

ABSTRACT Rapid identification and susceptibility testing results are of importance for the early appropriate therapy of bloodstream infections. The ePlex (GenMark Diagnostics) blood culture identification (BCID) panels are fully automated PCR-based assays designed to identify Gram-positive and Gram-negative bacteria, fungi, and bacterial resistance genes within 1.5 h from positive blood culture. Consecutive non-duplicate positive blood culture episodes were tested by the ePlex system prospectively. The choice of panel(s) (Gram-positive, Gram-negative, and/or fungal pathogens) was defined by Gram-stained microscopy of blood culture-positive bottles (BacT/Alert; bioMérieux). Results with the ePlex panels were compared to the identification results obtained by standard culture-based workflow. In total, 216 positive blood culture episodes were evaluable, yielding 263 identification results. The sensitivity/positive predictive value for detection by the ePlex panels of targeted cultured isolates were 97% and 99% for the Gram-positive panel and 99% and 96% for the Gram-negative panel, resulting in overall agreement rates of 96% and 94% for the Gram-positive and Gram-negative panel, respectively. All 26 samples with targeted resistance results were correctly detected by the ePlex panels. The ePlex panels provided highly accurate results and proved to be an excellent diagnostic tool for the rapid identification of pathogens causing bloodstream infections. The short time to results may be of added value for optimizing the clinical management of patients with sepsis.

scholarly journals Diagnosis of Bloodstream Infections in Children

2016 ◽  
Vol 54 (6) ◽  
pp. 1418-1424 ◽  
Author(s):  
Jennifer Dien Bard ◽  
Erin McElvania TeKippe
Keyword(s):  
Blood Culture ◽  
Clinical Microbiology ◽  
Bloodstream Infections ◽  
Clinical Microbiology Laboratory ◽  
Microbiology Laboratory ◽  
Diagnostic Approaches ◽  
Pediatric Populations ◽  
Culture Specimen

Identification of bloodstream infections is among the most critical tasks performed by the clinical microbiology laboratory. While the criteria for achieving an adequate blood culture specimen in adults have been well described, there is much more ambiguity in pediatric populations. This minireview focuses on the available pediatric literature pertaining to the collection of an optimal blood culture specimen, including timing, volume, and bottle selection, as well as rapid diagnostic approaches and their role in the management of pediatric bloodstream infections.

Sequence-Specific Targeting of Duplex DNA by Peptide Nucleic Acids via Triplex Strand Invasion

Methods ◽  
2001 ◽  
Vol 23 (2) ◽  
pp. 108-122 ◽  
Author(s):  
Vadim V. Demidov ◽  
Maxim D. Frank-Kamenetskii
Keyword(s):  
Nucleic Acids ◽  
Peptide Nucleic Acids ◽  
Duplex Dna ◽  
Specific Targeting ◽  
Strand Invasion

scholarly journals Rapid identification of Candida glabrata in Candida bloodstream infections

2007 ◽  
Vol 56 (12) ◽  
pp. 1639-1643 ◽  
Author(s):  
Nicholas Foster ◽  
Charlotte Symes ◽  
Richard Barton ◽  
Richard Hobson
Keyword(s):  
Blood Culture ◽  
Predictive Value ◽  
Candida Glabrata ◽  
Detection System ◽  
Bloodstream Infections ◽  
Rapid Identification ◽  
Resistant Organism ◽  
Blood Culture System ◽  
Aerobic Culture ◽  
Anaerobic Bottle

Candida species are the fourth most common cause of bloodstream infection (BSI) in the hospitalized patient. Candida glabrata is the most common non-Candida albicans Candida species in England and Wales with an attributed mortality of 48 %. C. glabrata is known to demonstrate reduced susceptibility to fluconazole, resulting in treatment failures when employing this agent for empirical treatment of Candida BSI. The first part of this study demonstrated a technique utilizing a blood culture system commonly used by many laboratories (BACTEC 9240 automated detection system) that reduced the time to identification of this potentially resistant organism by up to 72 h. A presumptive identification was achieved by observing a difference in the duration of incubation required before growth was detected automatically between Lytic Anaerobic and Plus Aerobic culture bottles. Secondly, experiments exploring the growth characteristics of C. glabrata in BACTEC blood culture bottles containing various media were carried out to explore possible reasons underpinning this clinical observation. The detection of yeast in the anaerobic bottle of a blood culture pair consisting of Lytic Anaerobic and Plus Aerobic in a BACTEC 9240 system was found to be highly predictive of the isolation of C. glabrata (positive predictive value 93.3 %, negative predictive value 98.3 %). The reason for this appeared to be a component of the Lytic Anaerobic blood culture medium enhancing the growth of C. glabrata in that medium.

scholarly journals 300. Pediatric Center Evaluation of the BioFire® Blood Culture Identification 2 Panel Versus the Original BioFire®FilmArray® Blood Culture Identification Panel for the Detection of Microorganisms and Resistance Markers in Positive Blood Cultures

2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S148-S149
Author(s):  
Kristina B Pierce ◽  
Rebecca Barr ◽  
Aubrie Hopper ◽  
Charlotte Bowerbank ◽  
Anne Shaw ◽  
...  
Keyword(s):  
Blood Culture ◽  
False Negative ◽  
Bloodstream Infections ◽  
Turnaround Time ◽  
Rapid Identification ◽  
Blood Cultures ◽  
Species Level ◽  
Genus Level ◽  
Antimicrobial Resistance Genes ◽  
Culture Identification

Abstract Background Studies show a rising annual incidence of severe sepsis, with bloodstream infections continuing to impact children. Rapid identification of causative agents and timely administration of targeted therapy can positively impact patient outcomes and improve antibiotic stewardship. The BioFire® Blood Culture Identification 2 (BCID2) Panel (BioFire Diagnostics, LLC), an updated version of the FDA-cleared BioFire® FilmArray® Blood Culture Identification (BCID) Panel, designed for use on positive blood cultures (PBCs), assesses 43 analytes, including 17 novel analytes (8 bacterial, 2 fungal, and 7 antimicrobial resistance genes), with a similar turnaround time. Methods De-identified residual PBCs for which clinician-ordered testing per standard of care (SoC) had been performed were enrolled and tested with an Investigation-Use-Only version of the BCID2 Panel. Only one positive bottle per patient was enrolled. Results of BCID2 and BCID were compared. Results 116 PBCs (48 aerobic and 68 anaerobic) were evaluated using the BioFire BCID2 Panel and results were compared to the BioFire BCID Panel. Of the 116 cases, 103 were positive on both the BioFire BCID2 Panel and the BioFire BCID Panel. Ten cases were negative on both tests. While the two panels showed 97% agreement, three cases were discrepant. Using culture (SoC) as the tiebreaker, two cases were false positive and one case was false negative on the BioFire BCID Panel. In all three cases, results from culture and the BioFire BCID2 Panel were in agreement. As expected, no organisms were detected on the BioFire BCID2 Panel in PBCs from 10% (12/116) of PBC bottles where culture identified only organisms that are not part of the panel menu. With the BioFire BCID2 Panel’s expanded platform, two cases identified as Enterobacteriaceae on the BioFire BCID Panel were identified to the genus level on the BioFire BCID2 Panel; 31 cases detected to the genus level on the BioFire BCID Panel were identified to the species level on the BioFire BCID2 Panel. Conclusion Overall, the BioFire BCID2 Panel performed well against the BioFire BCID Panel for identification of bloodstream pathogens and provided additional discrimination of some pathogens to the genus or species level. Data presented are from assays that have not been cleared or approved for diagnostic use. Disclosures All Authors: No reported disclosures

scholarly journals Epidemiology of clinically suspected and laboratory-confirmed bloodstream infections at a South African neonatal unit

2021 ◽  
Vol 15 (07) ◽  
pp. 943-952
Author(s):  
Angela Dramowski ◽  
Adrie Bekker ◽  
Mark Frederic Cotton ◽  
Andrew Christopher Whitelaw ◽  
Susan Coffin
Keyword(s):  
At Risk ◽  
Blood Culture ◽  
Fungal Pathogens ◽  
Case Fatality ◽  
Bloodstream Infections ◽  
Neonatal Unit ◽  
Risk Of Infection ◽  
Maternal Risk ◽  
Suspected Infection ◽  
Healthcare Associated

Introduction: Data from Africa reporting the epidemiology of infection in hospitalised neonates are limited. Methodology: A prospective study with convenience sampling was conducted to characterise neonates investigated with blood culture/s for suspected infection at a 132-bed neonatal unit in Cape Town, South Africa (1 February-31 October 2018). Enrolled neonates were classified as having proven bloodstream infection (BSI) (blood culture-positive with a pathogen) or presumed infection (clinically suspected but blood culture-negative) or as potentially at risk of infection (maternal risk factors at birth). Results: Of 1299 hospitalised neonates with >1 blood culture sampling episode, 712 (55%) were enrolled: 126 (17.7%) had proven BSI; 299 (42%) had presumed infection and 287 (40.3%) were potentially at risk of infection. Neonates with proven BSI had lower birth weight and higher rates of co-existing surgical conditions versus the presumed/potential infection groups (p < 0.001). Median onset of proven BSI versus presumed infection was at 8 (IQR = 5-13) and 1 (IQR = 0-5) days respectively (p < 0.001). Most proven BSI were healthcare-associated (114/126; 90.5%), with Klebsiella pneumoniae (80.6% extended-spectrum β-lactamase producers) and Staphylococcus aureus (66.7% methicillin-resistant) predominating. Mortality from proven BSI (34/126; 27%) was substantially higher than that observed in presumed (8/299; 2.7%) and potential infections (3/287; 1.0%) (p < 0.001). The odds of death from proven BSI was 3-fold higher for Gram-negatives than for Gram-positive/fungal pathogens (OR = 3.23; 95% CI = 1.17-8.92). Conclusions: Proven BSI episodes were predominantly healthcare-associated and associated with a high case fatality rate. Most neonates with presumed infection or at potential risk of infection had favourable 30-day outcomes.

scholarly journals Kinetic analysis of specificity of duplex DNA targeting by homopyrimidine peptide nucleic acids

1997 ◽  
Vol 72 (6) ◽  
pp. 2763-2769 ◽  
Author(s):  
V.V. Demidov ◽  
M.V. Yavnilovich ◽  
M.D. Frank-Kamenetskii
Keyword(s):  
Nucleic Acids ◽  
Kinetic Analysis ◽  
Peptide Nucleic Acids ◽  
Duplex Dna ◽  
Dna Targeting

scholarly journals Randomized Trial Evaluating Clinical Impact of RAPid IDentification and Susceptibility Testing for Gram-negative Bacteremia: RAPIDS-GN

2020 ◽  
Author(s):  
Ritu Banerjee ◽  
Lauren Komarow ◽  
Abinash Virk ◽  
Nipunie Rajapakse ◽  
Audrey N Schuetz ◽  
...  
Keyword(s):  
Blood Culture ◽  
Susceptibility Testing ◽  
Controlled Trial ◽  
Bloodstream Infections ◽  
Standard Of Care ◽  
Rapid Identification ◽  
Gram Negative ◽  
Soc Testing ◽  
Multicenter Randomized Controlled Trial ◽  
Gram Negative Bacteremia

Abstract Background Rapid blood culture diagnostics are of unclear benefit for patients with gram-negative bacilli (GNB) bloodstream infections (BSIs). We conducted a multicenter, randomized, controlled trial comparing outcomes of patients with GNB BSIs who had blood culture testing with standard-of-care (SOC) culture and antimicrobial susceptibility testing (AST) vs rapid organism identification (ID) and phenotypic AST using the Accelerate Pheno System (RAPID). Methods Patients with positive blood cultures with Gram stains showing GNB were randomized to SOC testing with antimicrobial stewardship (AS) review or RAPID with AS. The primary outcome was time to first antibiotic modification within 72 hours of randomization. Results Of 500 randomized patients, 448 were included (226 SOC, 222 RAPID). Mean (standard deviation) time to results was faster for RAPID than SOC for organism ID (2.7 [1.2] vs 11.7 [10.5] hours; P &lt; .001) and AST (13.5 [56] vs 44.9 [12.1] hours; P &lt; .001). Median (interquartile range [IQR]) time to first antibiotic modification was faster in the RAPID arm vs the SOC arm for overall antibiotics (8.6 [2.6–27.6] vs 14.9 [3.3–41.1] hours; P = .02) and gram-negative antibiotics (17.3 [4.9–72] vs 42.1 [10.1–72] hours; P &lt; .001). Median (IQR) time to antibiotic escalation was faster in the RAPID arm vs the SOC arm for antimicrobial-resistant BSIs (18.4 [5.8–72] vs 61.7 [30.4–72] hours; P = .01). There were no differences between the arms in patient outcomes. Conclusions Rapid organism ID and phenotypic AST led to faster changes in antibiotic therapy for gram-negative BSIs. Clinical Trials Registration NCT03218397.

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