Evaluation of real-time PCR and pyrosequencing for screening incubating blood culture bottles from adults with suspected bloodstream infection

McCann, Chase D.; Moore, Miranda S.; May, Larissa; McCarroll, Matthew G.; Jordan, Jeanne A.

doi:10.1016/j.diagmicrobio.2014.11.014

Cited by 12 publications

(14 citation statements)

References 26 publications

(33 reference statements)

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“…Serum samples were thawed on ice for prior to extraction peptide extraction based on a previously described non‐proteolytic procedure 27 . Briefly, different amounts of serum were combined with weak cation exchange (WCX) magnetic beads and WCX magnetic bead binding buffer into 0.2 ml eppendorf (EP) tubes before mixing and incubation for 0.5 h at RT.…”

Section: Methodsmentioning

confidence: 99%

Preliminary exploration on the serum biomarkers of bloodstream infection with carbapenem‐resistant Klebsiella pneumoniae based on mass spectrometry

Bao

Ding

et al. 2021

Clinical Laboratory Analysis

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Background Carbapenem‐resistant K. pneumoniae (CRKP) bloodstream infections (BSI) must be rapidly identified to improve patient survival rates. This study investigated a new mass spectrometry‐based method for improving the identification of CRKP BSI and explored potential biomarkers that could differentiate CRKP BSI from sensitive. Methods Mouse models of BSI were first established. MALDI‐TOF MS was then used to profile serum peptides in CRKP BSI versus normal samples before applying BioExplorer software to establish a diagnostic model to distinguish CRKP from normal. The diagnostic value of the model was then tested against 32 clinical CRKP BSI and 27 healthy serum samples. Finally, the identities of the polypeptides used to establish the diagnostic model were determined by secondary mass spectrometry. Results 107 peptide peaks were shared between the CRKP and normal groups, with 18 peaks found to be differentially expressed. Five highly expressed peptides in the CRKP group (m/z 1349.8, 2091.3, 2908.2, 4102.1, and 8129.5) were chosen to establish a diagnostic model. The accuracy, specificity and sensitivity of the model were determined as 79.66%, 81.48%, and 78.12%, respectively. Secondary mass spectrometry identified the Fibrinogen alpha chain (FGA), Inter‐alpha‐trypsin inhibitor heavy chain H4 (ITIH4) and Serum amyloid A‐2 protein (SAA2) as the source of the 5 serum peptides. Conclusions We successfully established a serum peptide‐based diagnostic model that distinguished clinical CRKP BSI samples from normal healthy controls. The application of MALDI‐TOF MS to measure serum peptides, therefore, represents a promising approach for early BSI diagnosis of BSI, especially for multidrug‐resistant bacteria where identification is urgent.

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Section: Methodsmentioning

confidence: 99%

Preliminary exploration on the serum biomarkers of bloodstream infection with carbapenem‐resistant Klebsiella pneumoniae based on mass spectrometry

Bao

Ding

et al. 2021

Clinical Laboratory Analysis

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“…The clinical signs and symptoms are almost indistinguishable between these 2 types of infection [ 3 ]. Hence, in the majority of these bloodstream-infected patients, the causative infectious agents cannot be identified due to the nonspecific signs and symptoms [ 4 ]. Therefore, it is customary to start empirical broad-spectrum antimicrobial therapy [ 5 ], but this contributes to the emergence of drug-resistant strains of bacteria and leads to increased medical costs.…”

Section: Introductionmentioning

confidence: 99%

Variation of Circulating Inflammatory Mediators in Staphylococcus aureus and Escherichia coli Bloodstream Infection

et al. 2016

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BackgroundThe aim of this study was to examine the behavior of circulating inflammatory mediators and to exclude gram-positive from gram-negative bloodstream infections. Results may be helpful in selection of optimal specific antibiotic therapies.Material/MethodsMice (25–27 g) were randomized to 3 groups infected with Staphylococcus aureus (S. aureus) ATCC 25923, Escherichia coli (E. coli) ATCC 25922, or phosphate-buffered saline (PBS). The white blood cell count (WBC) and the concentrations of serum C-reactive protein (CRP), procalcitonin (PCT), interleukin (IL)-1α, IL-1β, IL-6, IL-10, monocyte chemotactic protein-1 (MCP-1), and macrophage inflammatory protein-1α (MIP-1α) were detected in blood samples at different time intervals after intravenous tail injection.ResultsThe results showed that compared to the control mice, infected animals exhibited significantly higher levels of all mediators after bacterial infection. Moreover, compared to the mice that received S. aureus, animals with E. coli infection showed significantly greater increases in serum IL-1α, IL-1β, IL-6, MCP-1, and MIP-1α levels.ConclusionsThese results suggest that the use of the analyzed serum markers at an early stage of bloodstream infection may give useful information for the clinician to distinguish gram-negative from gram-positive infections.

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“…Pyrosequencing analysis is one of the techniques used to study microbial communities in numerous samples such as abomasal ulcers [14], blood cultures [15], drinking water distribution systems [16], and refrigerators [17]. The present work is a continuation from Abdul-Mutalib et al [9,18] who also applied the same technique to study microbial communities on cutting boards collected from several food premises in Seri Kembangan, Malaysia.…”

Section: Discussionmentioning

confidence: 96%

Prevalence of spoilage microorganism, Pseudomonas spp. on restaurants cutting boards collected in Seri Kembangan, Malaysia.

Na¹,

Osman²,

S³

et al. 2019

J Nutr Food Sci

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Some of the enzymes produced by Pseudomonas are heat Due to the tropical climate that promotes the growth of most microorganisms, the food service industry in Malaysia faces a lot of challenges in order to ensure quality foods are served to the customers. One of the bacteria that are associated with food spoilage is Pseudomonas spp., and the presence of these bacteria on cutting boards could cross-contaminate food items during food handling. This study performed pyrosequencing analysis to investigate the existence of microbial communities on cutting boards collected from restaurants in Seri Kembangan, Malaysia. Most of the samples were dominated by Flavobacteriales, Enterobacteriales, Lactobacillales, and Pseudomonadales. This study discovered that, all samples of cutting boards were contaminated with Pseudomonas spp. With the mean bacterial count of 1.4 × 10 6 cfu/cm 2 , 5.1 × 10 5 cfu/cm 2 and 5.6 × 10 6 cfu/cm 2 on samples collected from clean, moderately clean and less clean food premises, respectively. The study shows that the contamination level of Pseudomonas spp. was not significantly different in different grades of food premises. Regardless of the status and grades of the food premises, they have the same likelihood to introduce spoilage bacteria like Pseudomonas spp. from cutting board to food if they neglect correct food handling measures. Therefore, all food handlers need to prioritize safe food handling to avoid food contamination and spoilage. Abstract

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Evaluation of real-time PCR and pyrosequencing for screening incubating blood culture bottles from adults with suspected bloodstream infection

Cited by 12 publications

References 26 publications

Preliminary exploration on the serum biomarkers of bloodstream infection with carbapenem‐resistant Klebsiella pneumoniae based on mass spectrometry

Preliminary exploration on the serum biomarkers of bloodstream infection with carbapenem‐resistant Klebsiella pneumoniae based on mass spectrometry

Variation of Circulating Inflammatory Mediators in Staphylococcus aureus and Escherichia coli Bloodstream Infection

Prevalence of spoilage microorganism, Pseudomonas spp. on restaurants cutting boards collected in Seri Kembangan, Malaysia.

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