Detection of bacteria in blood by centrifugation and filtration

Bernhardt, Mat; Pennell, D R; Almer, Laurel S.; Schell, Ronald F.

doi:10.1128/jcm.29.3.422-425.1991

Cited by 22 publications

(14 citation statements)

References 24 publications

(20 reference statements)

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“…Filtered-based methods have been shown to offer good bacterial recovery rates in this study and in previous studies [11]. Nonetheless, filtration-based methods may not be applicable for processing a large number of samples with greater technicality and more time required per sample, which in turn heightens the risk of contamination.…”

Section: Discussionmentioning

confidence: 59%

“…To optimize the workflow from patient to pathogen ID and AST, a variety of recovery methods have been developed for isolating bacteria directly from blood. Some of these recovery methods include mechanical filtration, centrifugation, sedimentation, red blood cell lysis, chemical capture on surfaces or beads and microfluidic techniques [11][12][13][14][15][16][17][18]. Recovering bloodstream pathogens efficiently from the patient is vitally important in order to confidently rule in or rule out bacterial infection, to identify the causative agent and perform AST accurately and quickly.…”

Section: Introductionmentioning

confidence: 99%

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Improving the recovery and detection of bloodstream pathogens from blood culture

Falconer

Hammond

Gillespie

2020

Journal of Medical Microbiology

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Introduction. Bloodstream infections (BSI) are growing in incidence and present a serious health threat. Most patients wait up to 48 h before microbiological cultures can confirm a diagnosis. Low numbers of circulating bacteria in patients with BSI mean we need to develop new methods and optimize current methods to facilitate efficient recovery of bacteria from the bloodstream. This will allow detection of positive blood cultures in a more clinically useful timeframe. Many bacterial blood recovery methods are available and usually include a combination of techniques such as centrifugation, filtration, serum separation or lysis treatment. Here, we evaluate nine different bacteria recovery methods performed directly from blood culture. Aim. We sought to identify a bacterial recovery method that would allow for a cost-effective and efficient recovery of common BSI pathogens directly from blood culture. Methods. Simulated E. coli ATCC 25922 blood culture was used as a model system to evaluate nine different bacteria recovery methods. Each method was assessed on recovery yield, cost, hands-on time, risk of contamination and ease of use. The highest scoring recovery method was further evaluated using simulated blood cultures spiked with seven of the most frequently occurring bloodstream pathogens. The recovery yield was calculated based on c.f.u. count before and after each recovery method. Independent t-tests were performed to determine if the recovery methods evaluated were significantly different based on c.f.u. ml−1 log recovery. Results. All nine methods evaluated successfully recovered E. coli ATCC 25922 from simulated blood cultures although the bacterial yield differed significantly. The MALDI-TOF intact cell method offered the poorest recovery with a mean loss of 2.94±0.37 log c.f.u. ml−1. In contrast, a method developed by Bio-Rad achieved the greatest bacterial yield with a mean bacteria loss of 0.27±0.013 log c.f.u. ml−1. Overall, a low-speed serum-separation method was demonstrated to be the most efficient method in terms of time, cost and recovery efficiency and successfully recovered seven of the most frequent BSI pathogens with a mean bacteria loss of 0.717±0.18 log c.f.u. ml−1. Conclusion. The efficiency of bacterial recovery can vary significantly between different methods and thereby can have a critical impact on downstream analysis. The low-speed serum-separation method offered a simple and effective means of recovering common BSI pathogens from blood culture and will be further investigated for use in the rapid detection of bacteraemia and susceptibility testing in clinical practice.

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Section: Discussionmentioning

confidence: 59%

Section: Introductionmentioning

confidence: 99%

Improving the recovery and detection of bloodstream pathogens from blood culture

Falconer

Hammond

Gillespie

2020

Journal of Medical Microbiology

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“…Fresh whole human blood was then seeded with a selected dilution to obtain a final concentration of 1 to 50 microorganisms per ml of blood. This inoculum was chosen with the aim of simulating a low-grade bacteremia (1,10). Once it was seeded, the blood was incubated for 2 h at 35 Ϯ 2ЊC to allow phagocytosis of the bacteria (11,21,27), and then the blood sample was processed by the blood culture technique that was to be tested.…”

Section: Methodsmentioning

confidence: 99%

“…Moreover, they may present problems such as contamination and variability in the efficiency of the recovery of enterobacteria, Staphylococcus spp., Streptococcus spp., anaerobic bacteria, yeasts, and Mycobacterium spp. (1,26,31).…”

mentioning

confidence: 99%

Evaluation and comparison of different blood culture techniques for bacteriological isolation of Salmonella typhi and Brucella abortus

Gaviria-Ruiz¹,

Cardona‐Castro²

1995

J Clin Microbiol

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An experimental study was carried out to evaluate and compare various noncommercial methods of blood culture for the isolation of Salmonella typhi and Brucella abortus from fresh human blood samples that had been artificially inoculated with 1 to 50 microorganisms per ml of blood. The methods compared included the Ruiz-Castañeda blood culture, broth blood culture, leukocyte lysis and direct plating on agar (WBL-P), leukocyte lysis and filtration, Ficoll-Hypaque centrifugation and filtration, Ficoll-Hypaque centrifugation, and Ficoll-Hypaque centrifugation and leukocyte lysis methods. Results with the WBL-P technique showed that S. typhi was isolated in 18 h, and its recovery rate was 36.6% (calculated from the number of CFU recovered per milliliter versus the number inoculated). B. abortus was isolated in 48 h by the same technique, and its recovery rate was 48.8%. The isolation times for the other blood culture techniques were between 36 and 44 h for S. typhi and 66 h for B. abortus. The techniques which relied on filtering systems for the recovery of S. typhi and B. abortus performed poorly. The WBL-P technique for the isolation of S. typhi and B. abortus is faster than the other methods tested.

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“…The principle of the isolation is shown in Figure 6 as in the case of the OncoQuick separation kit. Additionally, this method is used to select bacterial cells from the sample for bacteremia diagnostics [179]. The employment of several media with different density allows us to fractionate the blood sample accurately and prevent fraction contamination by RBC and platelets.…”

Section: Density Gradient Centrifugationmentioning

confidence: 99%

Detection of Rare Objects by Flow Cytometry: Imaging, Cell Sorting, and Deep Learning Approaches

Voronin

Kozlova

Verkhovskii

et al. 2020

IJMS

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Flow cytometry nowadays is among the main working instruments in modern biology paving the way for clinics to provide early, quick, and reliable diagnostics of many blood-related diseases. The major problem for clinical applications is the detection of rare pathogenic objects in patient blood. These objects can be circulating tumor cells, very rare during the early stages of cancer development, various microorganisms and parasites in the blood during acute blood infections. All of these rare diagnostic objects can be detected and identified very rapidly to save a patient’s life. This review outlines the main techniques of visualization of rare objects in the blood flow, methods for extraction of such objects from the blood flow for further investigations and new approaches to identify the objects automatically with the modern deep learning methods.

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Detection of bacteria in blood by centrifugation and filtration

Cited by 22 publications

References 24 publications

Improving the recovery and detection of bloodstream pathogens from blood culture

Improving the recovery and detection of bloodstream pathogens from blood culture

Evaluation and comparison of different blood culture techniques for bacteriological isolation of Salmonella typhi and Brucella abortus

Detection of Rare Objects by Flow Cytometry: Imaging, Cell Sorting, and Deep Learning Approaches

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