Diagnosis Of Septic Patients Research Articles

Role of Glucose Variability in the Diagnosis of Septic Patients

Background: Glucose variability is an indicator and independent predictors of mortality and severity of sepsis in critically septic patients of intensive care unit. Objectives: This study evaluated the relationship of glucose variability in critically ill septic patients in relation with serum lactate. Method: It is a prospective observational study was conducted in the Intensive Care Unit (ICU) in Department of Anesthesia, Pain, Palliative Care and Intensive Care, Dhaka Medical College hospital, Dhaka over a period of one year in between 1st January- 31st December, 2015. Total 51 septic adult patients were included in the study according to the selection criteria. In this study, 8 consecutive capillary blood samples were taken with a periodic interval of 3 hours starting from admission. Mean and standard deviation (SD) blood glucose were computed to see the glucose variability and agreement done with serum lactate and severity of sepsis. All collected data were registered documented and analyzed in the statistical program Statistical Package for Social Science (SPSS) version 20.0. Result: Among total 51 patients, 52.9% study cases were of 4th decade with a mean age of 46±12 years had a mild female predominance. Glycemic variability was taken as >2SD. High glycemic variability observed in 70.6% cases. Good agreement observed in glucose variability with serum lactate by Kappa Statistics. Conclusion: Glucose variability shows the prediction of sepsis and it could be used as a useful alternative tool to predict sepsis and severity of sepsis. Bangladesh Crit Care J March 2022; 10 (1): 10-14

Early recognition of sepsis-induced coagulopathy using the C2PAC index: a ratio of soluble type C lectin-like receptor 2 (sCLEC-2) level and platelet count

C-type lectin-like receptor 2 (CLEC-2) is a platelet-activated receptor expressed on the surface of platelet membranes. Soluble CLEC-2 (sCLEC-2) has been receiving attention as a predictive marker for thrombotic predisposition. The present study examined the relationship between sCLEC-2 level and degree of coagulation disorder in septic patients. Seventy septic patients were divided into the sepsis-induced disseminated intravascular coagulation (DIC) (SID) group (n = 44) and non-SID group (n = 26). The sCLEC-2 levels were compared between the two groups. Because we suspected that the sCLEC-2 level was affected by the platelet count, we calculated the sCLEC-2/platelet count ratio (C2PAC index). We further divided septic patients into four groups using the Japanese Association for Acute Medicine (JAAM) DIC scoring system (DIC scores: 0–1, 2–3, 4–5, and 6–8). The C2PAC index was significantly higher in the SID group (2.6 ± 1.7) compared with the non-SID group (1.2 ± 0.5) (P < .001). The C2PAC indexes in the four JAAM DIC score groups were 0.9 ± 0.3, 1.1 ± 0.3, 1.7 ± 0.7, and 3.6 ± 1.0, respectively, and this index increased significantly as the DIC score increased (P < .001). According to the receiver-operating curve analysis, the area under the curve (AUC) and optimal cutoff value for the diagnosis of SID were 0.8051 and 1.4 (sensitivity, 75.0%; specificity, 76.9%), respectively. When the C2PAC index and D-dimer level, one of the main fibrinolytic markers, were selected as predictive markers for SID diagnosis in stepwise multiple logistic regression analysis, it was possible to diagnose SID with a high probability (AUC, 0.9528; sensitivity, 0.9545; specificity, 0.8846). The C2PAC index is a useful predictor of SID progression and diagnosis in septic patients.

Automatic continuous EEG signal analysis for diagnosis of delirium in patients with sepsis

ObjectiveIn critical care, continuous EEG (cEEG) monitoring is useful for delirium diagnosis. Although visual cEEG analysis is most commonly used, automatic cEEG analysis has shown promising results in small samples. Here we aimed to compare visual versus automatic cEEG analysis for delirium diagnosis in septic patients. MethodsWe obtained cEEG recordings from 102 septic patients who were scored for delirium six times daily. A total of 1252 cEEG blocks were visually analyzed, of which 805 blocks were also automatically analyzed. ResultsAutomatic cEEG analyses revealed that delirium was associated with 1) high mean global field power (p < 0.005), mainly driven by delta activity; 2) low average coherence across all electrode pairs and all frequencies (p < 0.01); 3) lack of intrahemispheric (fronto-temporal and temporo-occipital regions) and interhemispheric coherence (p < 0.05); and 4) lack of cEEG reactivity (p < 0.005). Classification accuracy was assessed by receiver operating characteristic (ROC) curve analysis, revealing a slightly higher area under the curve for visual analysis (0.88) than automatic analysis (0.74) (p < 0.05). ConclusionsAutomatic cEEG analysis is a useful supplement to visual analysis, and provides additional cEEG diagnostic classifiers. SignificanceAutomatic cEEG analysis provides useful information in septic patients.

Increased Levels of Histone in Human Plasma in Septic Patients with DIC

<Background and Methods>Sepsis is a life-threatening condition that is characterized by a whole-body inflammatory state (called a systemic inflammatory response syndrome, SIRS). Histones are essential for packing DNA into the cell nucleus and also play role in regulating gene expression but the new study (Jun Xu et al, Nature Medicine 15. 1318, 2009) have shown histones released into blood stream at the onset of sepsis can have devastating effects (endothelial damage and organ failure etc). However, plasma levels of histone are unclear in septic patients with DIC. Therefore, We developed a sandwich ELISA to quantify serum or plasma histone H3 levels and measured plasma levels of histone H3 in septic patients with DIC (n=30).<Results>DIC was diagnosed on the Japanese Ministry of Health and Welfare (JMHW) DIC diagnostic criteria. The thrombin antithrombin complexes (TAT) levels were higher in septic patients with DIC as reported by others. The working range of this ELISA was 3-1000 ng/ml. In healthy volunteers, plasma levels of histone (H3) were not detectable. We detected high levels of extracellular hisotone (H3) in the plasma of all septic patients with DIC by sandwich ELISA. Mean plasma levels of hisotone (H3) were 70.5 ± 63.2 (12.9-336.5) ng/ml in septic patients with DIC. However, plasma levels of extracellular histone (H3) were negative in acute promyelocytic leukemia (APL)-induced DIC (n=5). Also, Plasma levels of histone were 10 ng/ml or less by sandwich ELISA in septic patients without DIC (n=8).<Conclusion>These results suggest that assess of extracellular hisotones in the plasma may be helpful in the making the diagnosis in septic patients with DIC. Also, it appears that extracellular hisotones in the plasma may contribute to develop DIC in septic patients. DisclosuresNo relevant conflicts of interest to declare.

Increased Levels of PTX3 in Human Plasma in Septic Patients with DIC

Abstract 4347Sepsis is a life-threatening condition that is characterized by a whole-body inflammatory state (called a systemic inflammatory response syndrome, SIRS). Long pentraxin PTX3 is an inflammatory mediator and a component of the humoral arm of innate immunity produced by neutrophils, macrophages, myeloid dendritic and endothelial cells. During sepsis a massive inflammatory activation and coagulation/fibrinolysis dysfunction occur. However, little is known about PTX3 in septic patients with DIC. Therefore, we measured PTX3s in the plasma from septic patients with DIC (n=20). Also, we investigated PTX3 in the plasma from septic patients without DIC (n=8) and acute promyelocytic leukemia (APL)-induced DIC (n=5). PTX3 in the plasma from human were measured using enzyme-linked immunosorbent assay (ELISA) kits (Perseus Proteomics Inc, Japan).The thrombin antithrombin complexes (TAT) levels were higher in both DIC patients as reported by others. We detected high levels of PTX3 in the plasma of all septic patients with DIC. Also, plasma levels of PTX3 were positive in septic patients without DIC. However, plasma levels of PTX3 were significantly higher in septic patients with DIC than in septic patients without DIC. Plasma levels of PTX3 ware significantly correlated with severity of septic DIC (platelet count and TAT level). We did not find any difference plasma levels of PTX3 in the APL patients with DIC. Moreover, high-mobility group box 1 (HMGB1) concentrations in the human plasma were determined using ELISA kit (Shino-Test, Japan). Plasma levels of HMBG1 were positive for 14 cases in 20 septic patients with DIC. Also, we did not find any difference plasma levels of HMGB1 in the APL patients with DIC.These results suggest that assess of PTX3 and HMBG1 in the plasma may be helpful in the making the diagnosis in septic patients with DIC. However, PTX3 seemed to be a better biomarker than HMGB1. It appears that PTX3 may contribute to the severity of septic DIC. Disclosures:No relevant conflicts of interest to declare.

Increased Levels of Histone In Human Plasma In Septic Patients with DIC.

AbstractAbstract 1422Sepsis is a life-threatening condition that is characterized by a whole-body inflammatory state (called a systemic inflammatory response syndrome, SIRS). Histones are essential for packing DNA into the cell nucleus and also play role in regulating gene expression but the new study (Jun Xu et al, Nature Medicine 15. 1318, 2009) have shown histones released into blood stream at the onset of sepsis can have devastating effects (endothelial damage and organ failure etc). However, about extracellular histones in the plasma from human, they have shown only one patient's data. Therefore, we measured extracellular hisotones in the plasma from septic patients with DIC (n=20). Also, we investigated extracellular hisotones in the plasma from septic patients without DIC (n=5). Extracellular histones in the plasma from human were examined by western blot analysis for histone H3 (H3).The thrombin antithrombin complexes (TAT) levels were higher in septic patients with DIC as reported by others. We detected high levels of extracellular hisotones (H3) in the plasma of all septic patients with DIC by western blotting. However, we did not find any extracellular histones (H3) in the plasma of fiver septic patients without DIC by western blotting. Moreover, high-mobility group box 1 (HMGB1) concentrations in the human plasma were determined using ELISA kit (Shino-Test, Japan). Plasma levels of HMBG1 were negative for four cases in 20 septic patients with DIC. Also, Plasma levels of HMBG1 were negative in septic patients without DIC.These results suggest that assess of extracellular hisotones in the plasma may be helpful in the making the diagnosis in septic patients with DIC. It appears that extracellular hisotones in the plasma may contribute to develop DIC in patients with sepsis. Disclosures:No relevant conflicts of interest to declare.

LightCycler SeptiFast in early diagnosis of sepsis: our experience

The conventional sepsis diagnosis, using the cultivation technique, needs 24 hours for bacterial identification and 36 hours for fungal. The use of empiric therapy makes the growth of bacteria and fungi slower or may yield negative findings in many cases of septic shock. The molecular technique can contribute to a more rapid and specific diagnosis in septic patients. SeptiFast detects 26 bacterial and fungal species DNAs, using the PCR in real time and giving results after 6 hours. This is important for de-escalation therapy and beginning of appropriate antibiotic treatment. The aim of this study is to evaluate the sensibility and the specificity of the SeptiFast test versus traditional diagnosis.

Quantification of Low-Picomolar Concentrations of TNF-α in Serum Using the Dual-Network Microfluidic ELISA Platform

For both research and diagnostic purposes, the ability to detect low levels of proteins in a cost- and time-effective manner is essential. In this study, the cytokine TNF-alpha (tumor necrosis factor-alpha), a widely used protein indicator of inflammatory response, was chosen to demonstrate the ability of the dual-network microfluidic ELISA (enzyme-linked immunosorbent assay) platform developed by the authors to rapidly quantify low concentrations of this biomarker in serum. Through the optimization of several experimental parameters, the system was shown to meet the requirements for fundamental and applied studies, while also being relevant for challenging clinical applications such as the diagnosis of septic patients. A sensitivity of 45 pg/mL (2.6 pM) in both culture medium and serum, with inter- and intravariations of less than 15%, was attained for the quantification of human TNF-alpha to a concentration of up to 500 pg/mL. The overall time for completion of the assay in eight parallel reactions was less than 1 h.

A multiplex real-time PCR assay for rapid detection and differentiation of 25 bacterial and fungal pathogens from whole blood samples

Early detection of bloodstream infections (BSI) is crucial in the clinical setting. Blood culture remains the gold standard for diagnosing BSI. Molecular diagnostic tools can contribute to a more rapid diagnosis in septic patients. Here, a multiplex real-time PCR-based assay for rapid detection of 25 clinically important pathogens directly from whole blood in <6 h is presented. Minimal analytical sensitivity was determined by hit rate analysis from 20 independent experiments. At a concentration of 3 CFU/ml a hit rate of 50% was obtained for E. aerogenes and 100% for S. marcescens, E. coli, P. mirabilis, P. aeruginosa, and A. fumigatus. The hit rate for C. glabrata was 75% at 30 CFU/ml. Comparing PCR identification results with conventional microbiology for 1,548 clinical isolates yielded an overall specificity of 98.8%. The analytical specificity in 102 healthy blood donors was 100%. Although further evaluation is warranted, our assay holds promise for more rapid pathogen identification in clinical sepsis.

Relationship between Procalcitonin Levels and Severity of DIC Induced by Bacterial Infection.

<Background and Methods> In the past few years, structural and functional studies have demonstrated an increasingly tight interplay between the coagulation and inflammatory systems. These studies support an important role of systemic inflammatory response syndrome (SIRS) in the development of disseminated intravascular coagulation (DIC) in these critically ill patients. Although PCT is important clinical marker, it role in the DIC process is unkown. We measured serum levels of PCT in septic patients with DIC (n=15) and acute promyelocytic leukemia (APL)-induced DIC (n=5). Serum levels of PCT were determined by the EIA (BIOMERIEMX, France).<Results> The thrombin antithrombin complexes (TAT) levels were higher in both DIC patients as reported by others. ?Levels of PCT elevated in all patients with bacterial infection. In the septic patients with DIC, we found significant elevations of serum PCT levels (97.3±72.3 ng/ml) as compared with septic patients with non overt DIC (30.5±42.4 ng/ml). All the patients that levels of PCT increased than 100 ng/ml died. However, we did not find any difference serum levels of PCT in the APL and EB virus infected patients with DIC.<Conclusion> These results suggest that assess of thrombin generation capacity may be helpful in the making the diagnosis in septic patients with DIC. It appears that serum levels of PCT may contribute to the severity of DIC induced bacterial infection.

Increased Thrombin Generation Capacity in Septic Patients with DIC.

Variety clinical conditions may cause systemic activation of coagulation, ranging from insignificant laboratory changes to severe disseminated intravascular coagulation (DIC). DIC consists of widespread systemic activation of coagulation, resulting in diffuse fibrin deposition in small and midsize vessels. However, little is known about thrombin generation capacity in patients with DIC. To investigate the thrombin generation capacity, we measured thrombin generation in septic patients with DIC (n=20) and acute promyelocytic leukemia (APL)-induced DIC (n=5). Thrombin generation was determined by the Throbogram-Thrombinoscope assay (Thermo Electron Corporation, Netherlands). The analyzed TG parameters ware the peak of thrombin activity (Peak) and endogenous thrombin potential (ETP).The thrombin antithrombin complexes (TAT) levels were higher in both DIC patients as reported by others. In the septic patients with DIC, we found significant elevations in peak of thrombin activity and ETP as compared with normal controls (determined in 17 healthy males and 14 healthy female). However, the peak of thrombin activity and ETP levels were severely decreased by 60% n the APL patients with DIC. There was slightly correlation between the ETP and TAT levels in septic patients with DIC. Also, there was correlation between the ETP and bleeding tendency in APL patients with DIC.These results suggest that assess of thrombin generation capacity may be helpful in the making the diagnosis in septic patients with DIC. It appears that assess of ETP may contribute to evaluate bleeding tendency in APL patients with DIC.Sepsis with DICAPL with DICControls (n=31)Peak (nM)325.3 ± 55.5*88.2 ± 47.2*281.2 ± 40.9ETP (nM.min)1 ± 326.1*652.1 ± 274.5*1469.2 ± 227.3TAT (μg/ml)28.4 ± 17.5*22.5 ± 15.3*<3*(P<0.05) significantly different controls

How To Optimize the Detection of the Left Shift with the Instrument COULTER LH750®.

The Coulter LH750® is a last generation automated hematology analyzer. In 1999 we evaluated the optimization of immature granulocytes/bands (IG/B) detection in the Coulter GEN*S (Blood Vol.94 n° 10 sup.1, 1999 page 45 b), in which we observed that the Mean Volume of the Neutrophil population (M NE V) and the standard deviation of the neutrophil volume (SD NE V) were the most useful parameters to improve the detection of the presence of IG/B. Recently the utility of SD NE V has been proposed in the diagnosis of septic patients.(ISLH 05). The purpose of this study is to confirm that the NE morphometric parameters in the LH750 have the same values and cut-off as the GEN*S to discriminate the cases with lefts shift and if it is possible to create rules/flags that use these parameters to allow the automated detection of IG/B with cut-off levels specific to the origin (emergency room, outpatients, intensive care etc.) of the patient sample.In addition we investigated if the parameters are useful when taking into account not only the % of IG/B, but also their absolute number as recommended by the NCCLS. The overall intent being to optimize the efficiency of our laboratory in the detection of IG/B by decreasing the number of smears, but without increasing the number of false negatives. We have selected from the routine samples of a 2000 beds hospital, 340 consecutive cases on the LH750® that triggered the bands /Immature granulocytes flags, Immature NE 1 (NE1) or Immature NE 2 (NE2). We considered the samples with more than 0,9 IG/B (103/mL) as positive. Blood smears were performed on all the positive samples (340) and a WBC differential was performed according the NCCLS protocol (H 20A). The percentage and absolute number of IG/B were collected. The statistical analysis was performed using Medcalc® version 8.1.0.0. A ROC Curve and the cut-off for the detection of positive samples with immature IG/B using the SD NE V and M NE V was calculated.RESULTS: For the flag NE1 the best parameter to select the postive cases with left shift was the SD NE V with a cut-off of >24,2. (although this flag is actually defined for a moderate left shift). For the flag NE2 the best parameter to select the postive cases with left shift was the M NE V with a cut-off of > 161.CONCLUSION: Before the study performed in 1999 we validated the Imm NE1 and Imm NE2 flags according to the NE% and NE# or the WBC values, that statistically demonstrated limited utility. Parameters which permitted to optimize the detection of IG/B, were the Neutrophile morphological parameters; M NE V and SD NE V. The values of the LH 750 are slightly different to those of the GEN*S. We will subsequently study their utility and advantages compared to the IG/B counting in the detection of bacterial infections and sepsis due to better reproducibility.