Effect of dilution on sedimentational separation of bacteria from blood

Anderson, Clifton M.; Pitt, William G.

doi:10.1002/btpr.3056

Cited by 3 publications

(1 citation statement)

References 40 publications

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“…In addition to direct whole blood lysis, our present study also examined doing blood-lysis filtration to collect bacteria from plasma from whole blood spun upon a previously described centrifugal-sedimentation device [10,[33][34][35][36][37]. A lysis solution is often needed for the plasma recovered after spinning on this device because the device does not separate all RBCs from the resulting plasma.…”

Section: Introductionmentioning

confidence: 99%

Bacterial Extraction and Concentration from Blood through Filtration Processes

Pitt

2023

AJBSR

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Blood stream infections (BSIs) are challenging to rapidly and precisely diagnose. Current clinical diagnostic methods for BSI identification require culturing the blood sample for many hours prior to identifying the bacteria and possible antibiotic resistance. Removing the culturing step from the bacterial identification process of a BSI provides a significant reduction in the processing time, but shifts the rate-limiting step from the growth time to the concentration procedure, since CFU levels in clinical blood samples can be as low as 10 CFU/mL in blood. This study developed a novel whole blood filtration method to concentrate bacteria from a BSI without culturing the blood or bacteria. Filtering whole blood achieved 100% bacterial removal from 5mL of whole blood in ~90 s, but the bacteria were difficult to remove from the filter. Bacterial removal from the filter after blood filtration was also investigated. At high laboratory loadings of bacteria into freshly collected human blood, a blood lysis solution of 3% Tween 80 followed by a 3% Pluronic F108 backflush solution achieved 80% removal of the bacteria from the filter. This is a major step in collection of bacteria directly from blood for rapid diagnostics without a culture amplification step and with a single backflush.

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Section: Introductionmentioning

confidence: 99%

Bacterial Extraction and Concentration from Blood through Filtration Processes

Pitt

2023

AJBSR

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Culture-free Rapid Isolation and Detection of Bacteria from Whole Blood at Clinically Relevant Concentrations

Miguélez,

Osaid,

Larsson

et al. 2024

Preprint

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Rapid detection and antibiotic susceptibility profiling of bacterial agents in the blood of sepsis patients are crucial for determining appropriate treatment. The low bacteria concentrations and high abundance of blood cells currently necessitate culture-based diagnostic methods, which can take several days. Here, we introduce a method to isolate bacteria from whole blood with high separation power by smart centrifugation, followed by detection through microscopy in microfluidic traps within 2 h without the need for blood culture. We detected E. coli, K. pneumonia, or E. faecalis from spiked samples of healthy human donor blood at clinically relevant concentrations as low as 9, 7 and 32 colony-forming units per ml of blood, respectively. This rapid isolation of living bacteria from blood at clinically relevant concentrations opens possibilities for rapid phenotypic antibiotic susceptibility testing for bloodstream infections without blood culture.

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Two-phase analysis of blood in microchannel architecture on plasma separation ability with dimensional variance

Khaliq,

Lee,

Kamran

et al. 2024

HFF

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Purpose This paper aims to study the effects of varying inlet channel angle in a novel microfluidic architecture blood plasma separation ability over range of hematocrit values (5–45%) at multiple flowrates. Design/methodology/approach CAD designs for both micro architectures were designed in SOILWORKS. In the second step, these designs were imported into ANSYS to perform where meshing, model selection, defining blood as two-phase material and boundary conditions are performed. Findings Separation efficiency values close to 100% with diluted blood and 65.2% with whole blood were observed. Straight channel inlet design has significantly better performance at high hematocrit levels, whereas at lower hematocrit levels, both designs had almost same outcome. Furthermore, lower flowrates have shown the highest separation efficiency for lower hematocrit levels, whereas at higher hematocrit percentages, higher flowrates have shown better separation effects for both designs. Furthermore, trends obtained for flow ratio and flowrates against separation efficiency are demonstrated. Research limitations/implications This study is based on blood modeled as two-phase flow, with the phases consisting of blood plasma as primary phase and red blood cells as secondary particulate phase. Practical implications Implications of this study are far reaching for point-of-care health-care systems. A practical system of this numerical study can provide a microchannel device which take very small amount of blood sample to separate it into constituents which can be coupled with detection module to detect a particular disease for which it is designed for. This microsystem can be very beneficial for remote areas where a large hospital facility is far away. Originality/value This study has carried out a detailed analysis on the ability of a novel microchannel architecture to separate blood plasma from other blood constituents. Inlet channel angle variation effects are observed over a range of hematocrit percentages. These trends are further investigated for three different flowrates to assess the microchannel design behavior.

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Effect of dilution on sedimentational separation of bacteria from blood

Cited by 3 publications

References 40 publications

Bacterial Extraction and Concentration from Blood through Filtration Processes

Bacterial Extraction and Concentration from Blood through Filtration Processes

Culture-free Rapid Isolation and Detection of Bacteria from Whole Blood at Clinically Relevant Concentrations

Two-phase analysis of blood in microchannel architecture on plasma separation ability with dimensional variance

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