Diagnosis of bloodstream infections from positive blood cultures and directly from blood samples: recent developments in molecular approaches

Peker, Nilay; Couto, Natacha; Sinha, Bhanu; Rossen, John W. A.

doi:10.1016/j.cmi.2018.05.007

Cited by 163 publications

(146 citation statements)

References 73 publications

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“…Early pathogen identification and determination of resistance are essential to successful treatment outcome in patients with bacteremia and sepsis (29)(30)(31). Although direct detection of pathogens from whole blood has been realized on a limited basis in the United States and Europe, because of many challenges with their performance and interpretation, and the costs associated with implementation, blood cultures remain the major diagnostic method for detection of bloodstream infections (31,32).…”

Section: Discussionmentioning

confidence: 99%

Clinical Performance of the Novel GenMark Dx ePlex Blood Culture ID Gram-Positive Panel

Carroll

Reid²,

Thornberg³

et al. 2020

J Clin Microbiol

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Rapid identification from positive blood cultures is standard of care (SOC) in many clinical microbiology laboratories. The GenMark Dx ePlex Blood Culture Identification Gram-Positive (BCID-GP) Panel is a multiplex nucleic acid amplification assay based on competitive DNA hybridization and electrochemical detection using eSensor technology. This multicenter study compared the investigational-use-only (IUO) BCID-GP Panel to other methods of identification of 20 Gram-positive bacteria, four antimicrobial resistance genes, and both Pan Candida and Pan Gram-Negative targets that are unique to the BCID-GP Panel. Ten microbiology laboratories throughout the United States collected residual, deidentified positive blood culture samples for analysis. Five laboratories tested both clinical and contrived samples with the BCID-GP Panel. Comparator identification methods included each laboratory’s SOC, which included matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) and automated identification systems as well as targeted PCR/analytically validated real-time PCR (qPCR) with bidirectional sequencing. A total of 2,342 evaluable samples (1,777 clinical and 565 contrived) were tested with the BCID-GP Panel. The overall sample accuracy for on-panel organisms was 89% before resolution of discordant results. For pathogenic Gram-positive targets (Bacillus cereus group, Enterococcus spp., Enterococcus faecalis, Enterococcus faecium, Staphylococcus spp., Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus lugdunensis, Listeria spp., Listeria monocytogenes, Streptococcus spp., Streptococcus agalactiae, Streptococcus anginosus group, Streptococcus pneumoniae, and Streptococcus pyogenes), positive percent agreement (PPA) and negative percent agreement (NPA) ranged from 93.1% to 100% and 98.8% to 100%, respectively. For contamination rule-out targets (Bacillus subtilis group, Corynebacterium, Cutibacterium acnes, Lactobacillus, and Micrococcus), PPA and NPA ranged from 84.5% to 100% and 99.9% to 100%, respectively. Positive percent agreement and NPA for the Pan Candida and Pan Gram-Negative targets were 92.4% and 95.7% for the former and 99.9% and 99.6% for the latter. The PPAs for resistance markers were as follows: mecA, 97.2%; mecC, 100%; vanA, 96.8%; and vanB, 100%. Negative percent agreement ranged from 96.6% to 100%. In conclusion, the ePlex BCID-GP Panel compares favorably to SOC and targeted molecular methods for the identification of 20 Gram-positive pathogens and four antimicrobial resistance genes in positive blood culture bottles. This panel detects a broad range of pathogens and mixed infections with yeast and Gram-negative organisms from the same positive blood culture bottle.

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Section: Discussionmentioning

confidence: 99%

Clinical Performance of the Novel GenMark Dx ePlex Blood Culture ID Gram-Positive Panel

Carroll

Reid²,

Thornberg³

et al. 2020

J Clin Microbiol

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“…These conventional screenings are usually slow, requiring an over-night culture (o/n), are labor-intense and require expertise for interpretation. The molecular methods, either DNA microarray-based and/or nucleic acid amplification-based remain the gold standard for the detection of CTX-Ms and other ESBLs [ 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 ]. Many of these assays correspond to home-made PCR assays [ 19 , 20 ], but several assays are commercially available for CTX-M detection alone [ 18 ], and others for detecting also the 5 main carbapenemases such as the Verigene ® system (Luminex Corporation, Austin, TX, USA) and the FilmArray ® system (bioMérieux, Marcy l’Etoile, France).…”

Section: Introductionmentioning

confidence: 99%

“…Many of these assays correspond to home-made PCR assays [ 19 , 20 ], but several assays are commercially available for CTX-M detection alone [ 18 ], and others for detecting also the 5 main carbapenemases such as the Verigene ® system (Luminex Corporation, Austin, TX, USA) and the FilmArray ® system (bioMérieux, Marcy l’Etoile, France). The two latter may be used directly from patients’ positive blood cultures [ 21 , 22 ]. However, these tests are usually slow (1–4 h), labor-intense, expensive, require specific equipment and trained staff in order to be implemented in any clinical laboratory.…”

Section: Introductionmentioning

confidence: 99%

A Lateral Flow Immunoassay for the Rapid Identification of CTX-M-Producing Enterobacterales from Culture Plates and Positive Blood Cultures

et al. 2020

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We have developed a lateral flow immunoassay (LFIA), named NG-Test CTX-M MULTI (NG-Test), to detect group 1, 2, 8, 9, 25 CTX-M producers from agar plates and from positive blood cultures in less than 15 min. The NG-Test was validated retrospectively on 113 well-characterized enterobacterial isolates, prospectively on 102 consecutively isolated ESBL-producers from the Bicêtre hospital and on 100 consecutive blood cultures positive with a gram-negative bacilli (GNB). The NG-Test was able to detect all CTX-M producers grown on the different agar plates used in clinical microbiology laboratories. No false positive nor negative results were observed. Among the 102 consecutive ESBL isolates, three hyper mucous isolates showed an incorrect migration leading to invalid results (no control band). Using an adapted protocol, the results could be validated. The NG-Test detected 99/102 ESBLs as being CTX-Ms. Three SHV producers were not detected. Among the 100 positive blood cultures with GNB tested 10/11 ESBL-producers were detected (8 CTX-M-15, 2 CTX-M-27). One SHV-2-producing-E. cloacae was missed. The NG-Test CTX-M MULTI showed 100% sensitivity and specificity with isolates cultured on agar plates and was able to detect 98% of the ESBL-producers identified in our clinical setting either from colonies or from positive blood cultures.

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“…Clinical diagnosis of BSI depends on traditional blood culture results, although bacterial identification by these conventional methods is time-consuming. Molecular diagnostic tests to identify the presence of methicillin-susceptible S. aureus (MSSA) and MRSA isolates directly in critical clinical specimens, such as blood cultures, are being used, providing results in 1–5 h without the requirement for bacterial growth onto agar media, with increasing frequency in guiding antimicrobial therapy for staphylococcal infections as part of antimicrobial stewardship programs [ 22 , 23 , 24 , 25 , 26 , 27 ]. In recent years, several molecular panels able to simultaneously detect many organisms, including MSSA/MRSA, in one sample have been developed for the diagnosis of infectious syndromes.…”

Section: Introductionmentioning

confidence: 99%

Rapid Detection of Methicillin-Resistant Staphylococcus aureus Directly from Blood for the Diagnosis of Bloodstream Infections: A Mini-Review

et al. 2020

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Staphylococcus aureus represents a major human pathogen able to cause a number of infections, especially bloodstream infections (BSI). Clinical use of methicillin has led to the emergence of methicillin-resistant S. aureus (MRSA) and MRSA-BSI have been reported to be associated with high morbidity and mortality. Clinical diagnosis of BSI is based on the results from blood culture that, although considered the gold standard method, is time-consuming. For this reason, rapid diagnostic tests to identify the presence of methicillin-susceptible S. aureus (MSSA) and MRSA isolates directly in blood cultures are being used with increasing frequency to rapidly commence targeted antimicrobial therapy, also in the light of antimicrobial stewardship efforts. Here, we review and report the most common rapid non-molecular and molecular methods currently available to detect the presence of MRSA directly from blood.

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Diagnosis of bloodstream infections from positive blood cultures and directly from blood samples: recent developments in molecular approaches

Cited by 163 publications

References 73 publications

Clinical Performance of the Novel GenMark Dx ePlex Blood Culture ID Gram-Positive Panel

Clinical Performance of the Novel GenMark Dx ePlex Blood Culture ID Gram-Positive Panel

A Lateral Flow Immunoassay for the Rapid Identification of CTX-M-Producing Enterobacterales from Culture Plates and Positive Blood Cultures

Rapid Detection of Methicillin-Resistant Staphylococcus aureus Directly from Blood for the Diagnosis of Bloodstream Infections: A Mini-Review

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