White Blood Cell Count Levels Research Articles

The inflammatory markers of multisystem inflammatory syndrome in children (MIS-C) and adolescents associated with COVID-19: A meta-analysis.

To conduct a systematic review and meta‐analysis to characterize inflammatory markers in comparisons of multisystem inflammatory syndrome in children (MIS‐C) versus severe/non‐severe COVID‐19, severe MIS‐C versus non‐severe MIS‐C, and among age groups of MIS‐C. Nine databases were searched for studies on inflammatory markers of MIS‐C. After quality checks, data were pooled using a fixed or random effects model. Inflammatory markers included white blood cell count (WBC) or leukocytes, absolute lymphocyte count (ALC), absolute neutrophil count (ANC), platelet count (PLT), C‐reactive protein (CRP), procalcitonin (PCT), ferritin, D‐dimer, lactate dehydrogenase (LDH), fibrinogen, and erythrocyte sedimentation rate (ESR) for comparisons by severity and age. Twenty‐one studies with 1735 participants yielded 787 MIS‐C patients. Compared to non‐severe COVID‐19 patients, MIS‐C patients had lower ALC and higher ANC, CRP, and D‐dimer levels. Compared to severe COVID‐19 patients, MIS‐C patients had lower LDH and PLT counts and higher ESR levels. Severe MIS‐C patients had higher levels of WBC, ANC, CRP, D‐dimer, and ferritin than non‐severe MIS‐C patients. For MIS‐C, younger children (0–5 years) had lower CRP and ferritin levels than middle‐aged/older children/adolescents. Measurement of inflammatory markers might assist clinicians in accurate evaluation and diagnosis of MIS‐C and the associated disorders.

Laboratory biomarkers of COVID-19 disease severity and outcome: Findings from a developing country.

AimTo identify laboratory biomarkers that predict disease severity and outcome among COVID-19 patients admitted to the Millennium COVID-19 Care Center in Ethiopia.MethodsA retrospective cohort study was conducted among 429 COVID-19 patients who were on follow up from July to October 2020. Data was described using frequency tables. Robust Poisson regression model was used to identify predictors of COVID-19 severity where adjusted relative risk (ARR), P-value and 95 CI for ARR were used to test significance. Binary Logistic regression model was used to assess the presence of statistically significant association between the explanatory variables and COVID-19 outcome where adjusted odds ratio (AOR), P-value and 95%CI for AOR were used for testing significance.ResultsAmong the 429 patients studied, 182 (42.4%) had Severe disease at admission and the rest 247 (57.6%) had Non-severe disease. Regarding disease outcome, 45 (10.5%) died and 384 (89.5%) were discharged alive. Age group (ARR = 1.779, 95%CI = 1.405–2.252, p-value <0.0001), Neutrophil to Lymphocyte ratio (NLR) (ARR = 4.769, 95%CI = 2.419–9.402 p-value <0.0001), Serum glutamic oxaloacetic transaminase (SGOT) (ARR = 1.358, 95%CI = 1.109–1.662 p-value = 0.003), Sodium (ARR = 1.321, 95%CI = 1.091–1.600 p-value = 0.004) and Potassium (ARR = 1.269, 95%CI = 1.059–1.521 p-value = 0.010) were found to be significant predictors of COVID-19 severity.The following factors were significantly associated with COVID-19 outcome; age group (AOR = 2.767, 95%CI = 1.099–6.067, p-value = 0.031), white blood cell count (WBC) (AOR = 4.253, 95%CI = 1.918–9.429, p-value = 0.0001) and sodium level (AOR = 3.435, 95%CI = 1.439–8.198, p-value = 0.005).ConclusionsAssessing and monitoring the laboratory markers of WBC, NLR, SGOT, sodium and potassium levels at the earliest stage of the disease could have a considerable role in halting disease progression and death.

Mortality Following Clostridioides difficile Infection in Europe: A Retrospective Multicenter Case-Control Study.

We aimed to describe the clinical presentation, treatment, outcome and report on factors associated with mortality over a 90-day period in Clostridioides difficile infection (CDI). Descriptive, univariate, and multivariate regression analyses were performed on data collected in a retrospective case-control study conducted in nine hospitals from seven European countries. A total of 624 patients were included, of which 415 were deceased (cases) and 209 were still alive 90 days after a CDI diagnosis (controls). The most common antibiotics used previously in both groups were β-lactams; previous exposure to fluoroquinolones was significantly (p = 0.0004) greater in deceased patients. Multivariate logistic regression showed that the factors independently related with death during CDI were older age, inadequate CDI therapy, cachexia, malignancy, Charlson Index, long-term care, elevated white blood cell count (WBC), C-reactive protein (CRP), bacteraemia, complications, and cognitive impairment. In addition, older age, higher levels of WBC, neutrophil, CRP or creatinine, the presence of malignancy, cognitive impairment, and complications were strongly correlated with shortening the time from CDI diagnosis to death. CDI prevention should be primarily focused on hospitalised elderly people receiving antibiotics. WBC, neutrophil count, CRP, creatinine, albumin and lactate levels should be tested in every hospitalised patient treated for CDI to assess the risk of a fatal outcome.

Baseline characteristics and changes of biomarkers in disease course predict prognosis of patients with COVID-19.

The outbreak of coronavirus disease (COVID-19) has brought great challenges to the world. The objectives of this study were to describe the baseline characteristics and changes of biomarkers of these COVID-19 patients and identify predictive value of the above markers for patient death. Using patient death as the observational endpoints, clinical data of inpatients in a special ward for COVID-19 in Wuhan, China were retrospectively collected. Univariate and multivariate Cox regression analyses were used to evaluate prognostic value of baseline characteristics and laboratory data changes. This study included clinical data of 75 patients. Age, c-reactive protein (CRP) and interleukin-6 levels were independent predictors of patient death. Survivors were characterized as having declining neutrophil counts, d-dimer, N-terminal pronatriuretic peptide, troponin I (TnI) and c-reactive protein levels, while counts of lymphocyte gradually came back. Non-survivors were characterized with increasing white blood cell counts (WBC) and neutrophil counts. Changes of WBC, TnI and interleukin-6 were also independently associated with patient death. Older age, baseline CRP and IL-6 levels may be used as meaningful predictors to identify patients with poor prognosis. Changes of biomarkers should be closely monitored in the management of patients with COVID-19, while constantly increasing levels of WBC, TnI and interleukin-6 in the disease course also predict patient death.

Markers of Chronic Inflammation in Overweight and Obese Individuals and the Role of Gender: A Cross-Sectional Study of a Large Cohort.

ObjectiveDuring the last decade, obesity has become an epidemic. As obesity is now considered a state of low-grade inflammation, the purpose of this study was to assess the prevalence of four common elements of inflammation, in individuals with increased BMI. These findings were compared to those of subjects with normal BMI. The effect of gender was also noted.MethodsData were collected from medical records of individuals examined at a screening center in Israel between the years 2000–2014. Cross-sectional analysis was carried out on 7526 men and 3219 women. White blood cell count (WBC); platelet (PLT) count; erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) were assessed in four BMI categories: normal, overweight, obese and morbidly obese.ResultsMean (SD) age of the study sample was 47.5 (9.7) and 46.7 (9.8) years for men and women, respectively. The prevalence of each inflammatory marker increased significantly when comparing abnormal to normal BMI (p<0.0001). The odds ratio (OR) of the prevalence of increased inflammatory markers was compared between subjects with overweight, obese and morbid obesity and subjects with normal BMI. This study showed that the higher the BMI, the higher the OR. For those in the morbid obesity group, the OR for the different inflammatory markers adjusting for age, diabetes mellitus hypertension and kidney function were as follows: WBC levels, 5.1 (2.9–8.7) and 4.7 (2.4–9.1) for men and women, respectively; PLT levels, 1.7 (0.3–8.5) and 2.0 (0.6–7.2) for men and women, respectively; ESR levels, 4.2 (3.2–5.4) and 4.6 (3.2–6.6) for men and women, respectively, and CRP levels, 13.4 (10.0–18.2) and 19.2 (12.9–28.6) for men and women, respectively.ConclusionInflammatory markers are significantly higher in subjects with abnormal compared to normal BMI. This difference was found to be greater in women than in men.

Effect of thoraco-laparoscopic esophagectomy on postoperative immune function of patients with esophageal carcinoma

To investigate the effect of thoraco-laparoscopic esophagectomy on postoperative immune function of patients with esophageal carcinoma. Eighty-one patients undergoing radical esophagectomy in our hospital between January, 2017 and December, 2019 were enrolled in this study.According to the surgical approach, the patients were divided into endoscopic group (41 cases) and open surgery (3 incisions) group (40 cases).The immunological indicators (CD3+, CD4+, CD8+, and CD4+/CD8+ratio) of the patients were analyzed using double antibody sandwich enzyme-linked immunosorbent assay at 1 day before the surgery and on days 1, 4 and 7 after the surgery.The plasma levels of interleukin 6(IL-6) and cortisol, β-endorphin (β-EP) level, white blood cell count (WBC) and C-reactive protein (CRP) levels of the patients were measured before and on days 1 and 7 after the operation. No death occurred in either of the group after the operation.On days 4 and 7 after the operation, CD3+, CD4+, CD8+, and CD4+/CD8+ratio were significantly higher in patients undergoing thoracolaparoscopic surgery than in those receiving open surgery (P < 0.05).In both groups, the levels of cortisol, β-EP, WBC, CRP and IL-6 measured on days 1 and 7 postoperatively were significantly different from those before the operation (P < 0.05).At all the indicated postoperative time points, all the measured indicators, with the exception of IL-6 levels on postoperative day 7, which were comparable between the two groups, were significantly higher in the open surgery group than in the endoscopic group (P < 0.05). Thoraco-laparoscopic resection of esophageal cancer can reduce postoperative secretion of proinflammatory factors, alleviate inflammatory responses, and promote the recovery of immune functions to accelerate postoperative recovery of the patients.

The clinical characteristics of coronavirus disease 2019 in mild and ordinary female patients

AIMS: the aim of study was to explore the clinical characteristics of coronavirus disease 2019 in mild and ordinary patients, providing guidance for clinical diagnosis and treatment.&#x0D; MATERIALS AND METHODS: Sixty-four patients with positive results of nucleic acid testing were divided into two groups, called the youth group and the older group, to be compared the differences of clinical characteristics.&#x0D; RESULTS: Sixty-nine cases showed normal or decrease in white blood cell count (WBC). One case decreased in lymphocyte count (LYMPH). Seven cases decreased in lymphocyte proportion (LYMPH%). Seven cases increased in C-reactive protein (CRP). Sixteen cases increased in alanine transaminase (ALT). Nineteen cases increased in aspartate transaminase (AST). One case increased in lactic dehydrogenase (LDH). Six cases decreased in creatine kinase (CK). Three cases increased in globulin (GLOB). Thirty-eight cases decreased in serum creatinine (CRE). Five cases decreased in blood urea nitrogen (BUN). The lung CT scanning results showed no statistic difference between two groups (p 0.05). The level of WBC, NEUT, LDH, BUN, potassium (K+) showed statistic difference between two groups (p ˂ 0.05).&#x0D; CONCLUSIONS: Age was a risk factor in COVID-19. In the course of future diagnosis and treatment, blood test might play an increasingly important role.

Amelioration of systemic inflammation in advanced chronic liver disease upon beta-blocker therapy translates into improved clinical outcomes

ObjectiveSystemic inflammation promotes the development of clinical events in patients with advanced chronic liver disease (ACLD). We assessed whether (1) non-selective beta blocker (NSBB) treatment initiation impacts biomarkers of systemic...

Application of procalcitonin, white blood cell count and neutrophil-to-lymphocyte ratio in the diagnosis of systemic lupus erythematosus with a bacterial infection.

Systemic lupus erythematosus (SLE) is an autoimmune disease. This study aims to analyze the value of procalcitonin (PCT), white blood cell count (WBC), and neutrophil-to-lymphocyte ratio (NLR) in the differential diagnosis of SLE with a bacterial infection. A total of 164 patients with SLE admitted to our hospital from January 2018 to December 2019 were selected as the research subjects. According to the results of bacterial culture, patients with SLE were divided into the bacterial infection group (46 cases) and the non-bacterial infection group (118 cases). Sixty healthy volunteers without the allergic disease, family history of tumors, or immunodeficiency in the same age group were selected as the control group during the same period. Fasting venous blood was collected in the morning to detect serum macrophage migration inhibitory factor (MIF), C-reactive protein (CRP), red blood cell distribution width (RDW), PCT, WBC, neutrophil count (NEUT) and lymphocyte count (LYMPH), and the NLR (NEUT/LYMPH) was calculated. CRP, PCT, WBC, and NLR levels were significantly higher in the bacterial infection group and the non-bacterial infection group than that in the control group, and which in the bacterial infection group is significantly higher than in the non-bacterial infection group (P<0.05). PCT, WBC and NLR were independent risk factors for SLE with bacterial infection (P<0.05). ROC curve analysis showed the areas under the curve of PCT, WBC, and NLR for the differential diagnosis of SLE with bacterial infection being 0.883, 0.669, and 0.624. Then, the sensitivities of 78.3%, 43.5%, and 69.6% were observed with the specificities of 98.3%, 91.5%, and 56.8%, respectively. The area under the curve of combined prediction was 0.919, with a sensitivity of 78.3%, and a specificity of 97.5%. These results showed that the value of the combined prediction was significantly higher than the single prediction of each indicator. And the levels of PCT, WBC, and NLR gradually decreased with the progress of treatment (P<0.05). The combined detection of PCT, WBC, and NLR levels shows high sensitivity and specificity for SLE with a bacterial infection, which can be used for early auxiliary diagnosis of SLE with a bacterial infection.

Progression of Inflammatory Biomarkers Early after Cardiovascular Implantable Electronic Device Placement

Objective: There is a lack of data in the current literature regarding the spontaneous progression of inflammatory biomarkers early after cardiovascular implantable electronic device (CIED) placement. In our study we aimed to detect C-reactive protein (CRP) and white blood cell count (WBC) trends in the 5 days following CIED implantation. Methods: We retrospectively investigated a total of 235 patients who received CIEDs between 2012 and 2016 in our heart center. The levels of CRP and WBC for one day before and for the following 5 days after the procedure were noted in 4 serial measurements. The longitudinal course of CRP and WBC and their association with various parameters were analyzed with linear mix model of repeated measurements. Results: Totally 235 patients were screened; 36 patients excluded due to lack of serial CRP and WBC measurements, 44 patients excluded due to occurrence of a factor that may potentially affect CRP and WBC level. Finally 153 patients with a median age of 67 (IQR 58-74), 41 (26.8%) of whom were female, were included in the study. CRP values indicated a significant trend and made a peak in the second estimation (between 48-72 hours) after the procedure (Beta[β]: 0.208; Standard Error [S.E]: 0.015; p&amp;lt; 0.001). Age and procedure duration longer than 1 hour emerged as leading factors for high level of CRP (β:0.004; S.E: 0.002; p=0.05 and β: 0.208; S.E: 0.015; p= 0.02, respectively). The relation between time and WBC count was not statically significant (β: -0.129; S.E: 0.06; p=0.06). Conclusion: Progression of CRP after CIED implantation demonstrates a significant trend. Age and procedure duration longer than 1 hour are leading factors for high levels of CRP. There is statistically non-significant relation between WBC levels and time in 5 days of procedure.

Biological predictors of third-line treatment outcomes in patients with advanced non-small cell lung cancer.

e21597 Background: The introduction of molecularly-guided targeted therapies and immunotherapy in routine oncology practice has led to significant survival increases in a subset of patients with advanced-stage non-small cell lung cancer (NSCLC). Nevertheless, the ultimate prognosis of these patients remains grim, and their overall therapeutic management challenging, particularly beyond the first-line treatment setting. We herein aimed to assess outcomes of third-line treatment in real-world patients with NSCLC, and to investigate potential correlations of clinicopathological and laboratory fearures with survival data. Methods: The medical records of 111 patients with advanced NSCLC, diagnosed and treated at the Oncology Unit of Sotiria Athens General Hospital between January 1, 2015 and December 31, 2018, were retrospectively reviewed. Eligible patients were those who had received more than two cycles of third-line treatment. Clinicopathological and laboratory features of patients, including baseline levels of complete blood count parameters evaluated immediately before administration of third-line therapy, were recorded and correlated with progression-free and overall survival. Results: The majority of patients had lung adenocarcinoma (54.1%) and stage IV disease (64,9%). Progression-free survival rates at 3 and 6 months were 59% and 20, respectively. Risk of disease progression was 35% lower in adenocarcinoma, as compared to other histological subtypes. Increased haematocrit and reduced white blood cell count levels were independently correlated with decreased risk of disease progression [HR (95% CI): 0,95(0,91-1,00); p = 0,044 and HR (95% CI):.1,05(1,00-1,11); p = 0,048, respectively]. Furthermore, higher white blood cell count and platelet count levels were independent predictors of decreased survival [HR (95% CI): 1,05 (1,01-1,1); p = 0,018 and HR (95% CI): 1,02(1-1,04); p = 0,018, respectively]. Conclusions: Haematocrit, white blood cell count and platelet count, recorded immediately before initiation of third-line therapy, may represent independent predictors of survival parameters in patients with advanced NSCLC.

Reactive increase of cerebrospinal fluid white blood cell count after lumbar puncture: Fact or fiction?

Lumbar puncture (LP) is commonly used in the diagnostic workup of neurological patients, often to exclude inflammatory diseases of the central nervous system. In clinical practice, an increase of white blood cell count (WBC) in the cerebrospinal fluid (CSF) after a LP is often assumed as reactive to the first puncture. Scientific evidence of this hypothesis, however, is lacking.Retrospective review of laboratory parameters was done by analyzing CSF of patients who had at least two LPs between 2012 and 2016 in a single center. Inclusion criteria were a normal CSF WBC in the first LP as well as absence of any underlying disease typically associated with increased CSF WBC.A total of 176 patients (age 57.0 ± 17.6) with 260 serial LPs were included. No significant effect on the CSF WBC (1.2 ± 1.1 vs 1.4 ± 1.4/μl, p = .17), lactat and protein level between consecutive punctures was found after a second LP. In the subgroup of 104 patients who had two LPs within ten days, only one (0.96%) showed a mild abnormal CSF WBC (9 leukocytes/μl) in the second LP.A raise of CSF WBC after LP is rare and not commonly found; therefore, it should lead to careful exclusion of other, especially inflammatory diseases. The needle size is important to minimize the trauma during LP and seems to have an influence on the rate of reactive increase of CSF WBC after LP.

Prognostic analysis of severe patients with bloodstream infection caused by Enterobacteriaceae bacteria

To investigate the prognostic factors of severe patients with bloodstream infection (BSI) caused by Enterobacteriaceae bacteria. Patients suffered from BSI caused by Enterobacteriaceae bacteria admitted to department of critical care medicine of Binzhou Medical University Hospital from October 2016 to October 2019 were enrolled. The information of gender, age, combined shock, acute physiology and chronic health evaluation II (APACHE II), sequential organ failure assessment (SOFA), sensitivity of initial antibiotics, as well as the baseline of procalcitonin (PCT), white blood cell count (WBC), platelet (PLT), albumin (ALB) were collected. The 72-hour PCT clearance rate (72 h PCTc) was calculated after 72 hours' treatment. According to the clinical outcome after 28 days, the patients were divided into recovery group and death group. The differences of clinical indicators between the two groups were compared, and then the statistical significant variables were further performed by Logistic regression to analyze the factors affecting the prognosis of patients. The receiver operating characteristic (ROC) curve was drawn to evaluate the predictive efficacy of the factors in severe BSI. A total of 86 patients were enrolled, among whom 54 cases recovered while 32 cases died, and the 28-day mortality was 37.2%. There was no significant difference in gender, age, sensitivity of initial antibiotics, baseline levels of PCT and WBC between two groups. In the death group, the shock incidence, APACHE II score, SOFA score were significantly higher than those in recovery group [shock incidence: 84.4% (27/32) vs. 46.3% (25/54), APACHE II: 24.94±7.65 vs. 17.02±6.57, SOFA: 11.00±3.27 vs. 6.30±2.65, all P < 0.01]; the PLT and ALB baseline levels, 72 h PCTc were significantly lower than those in recovery group [PLT (×109/L): 73.38±49.15 vs. 138.69±101.80, ALB (g/L): 25.47±5.91 vs. 28.59±4.53, 72 h PCTc: -44 (-170, 27)% vs. 63 (40, 77)%, all P < 0.01]. The above 6 variables were included in Logistic regression. The results showed that SOFA score was a risk factor for death in these patients [odds ratio (OR) = 1.930, P = 0.037], while 72 h PCTc and ALB were protective factors (OR values were 0.043, 0.783, P values were 0.008, 0.047). The SOFA, 72 h PCTc and ALB can be used to predict the prognosis of severe BSI, and the diagnostic value of the combination of three factors was the largest [area under the ROC curve (AUC) = 0.953, 95% confidence interval (95%CI) was 0.909-0.997], the sensitivity was 100%, and the specificity was 79.6%. Severe patients with BSI caused by Enterobacteriaceae bacteria had a high mortality. Higher SOFA score, and lower ALB and 72 h PCTc predicted the adverse outcome. The combination of the three factors has the greatest prognostic efficacy.

Fever and inflammatory markers do not predict infection in pediatric ventricular assist device recipient

ObjectiveWe sought to assess the relationship between serious infections and fever, white blood cell count (WBC), and C-reactive protein (CRP) levels in children on ventricular assist device (VAD). MethodsWe retrospectively reviewed all children who underwent VAD placement from 2006 to 2014 at Children's Hospital of Pittsburgh. Daily maximum temperature, maximum WBC, CRP levels, microbiologic data and antibiotic use were abstracted from the electronic health record. Serious infection was defined as bacteremia, mediastinitis and/or driveline infection. ResultsThirty-six consecutive patients underwent VAD placement with mean age of 7.1 years ±6.3 years (range 9 days −18.1 years). The median duration of VAD support was 24 days (1–102 days). Patients were on antibiotics for 58% of total VAD days. We found no drive line infections or mediastinitis and only 5 episodes of blood stream infections in 4 patients (11%) during 1167 total days. Fever was present on 23% of total VAD days and nearly all (35/36) patients had at least one day of fever. CRP was elevated in 298 of 318 (94%) of all samples tested. 53% (448/851) of the total WBC assessments were elevated. On detailed analysis of confirmed bloodstream infection, only 2 of the 5 episodes were associated with fever within 24 h of the positive culture. WBC and CRP levels (drawn within 24 h of positive culture) were normal in 2/5 (40%). ConclusionsDespite a low prevalence of serious infection in our pediatric VAD cohort, we found that fever, elevated WBC and CRP were common. This suggests that traditional markers of infection are unreliable. Fever, elevated WBC count and elevated CRP were poorly predictive of serious bacterial infection in our pediatric VAD cohort. Their presence alone should not be an indication for long-term antibiotics.

Diagnostic value of presepsin in predicting a complicated acute appendicitis

Aim: There is a growing body of evidence suggesting that the concentration of presepsin in serum was found to be increased in patients with sepsis compared to healthy individuals, but the clinical significance of presepsin in the diagnosis of complicated acute appendicitis remains unclear. The purpose of this study was investigation of the diagnostic value of presepsin in complicated acute appendicitis. Material and Methods: This clinical prospective single-center study was conducted between May and August 2018, and comprised 120 patients with definitive diagnosis of acute appendicitis and 30 individuals as control group. White blood cell count (WBC), C-reactive protein (CRP), neutrophil/lymphocyte ratio (NLR) and presepsin levels were measured from venous blood samples. A receiver operating characteristic (ROC) curve was constructed to assess sensitivity and specificity as well as optimal cut points for each presepsin to diagnose acute appendicitis.Results: Serum levels of CRP, WBC, NLR and presepsin were higher in complicated group compared with uncomplicated (p0.0001, p=0.01, p=0.034 and p0.0001, respectively). The time of hospitalization, the rate of postoperative infection, intraabdominal abscess and re-hospitalization were significantly higher in complicated group (p0.0001, p=0.0009, p=0.0073 and p=0.0073, respectively). Comparing the diagnostic test results with ROC analysis, variable WBC (AUC=0.646, p=0.0052), CRP (AUC=0.807, p0.001), NLR (AUC=0.620, p=0.0305) and presepsin (AUC=0.965, p0.001) were significant parameters in predicting the presence of complicated acute appendicitis. Cut-off points of CRP, WBC, NLR and presepsin were calculated as 13.5 mg/dL, 1.4 x103/uL, 4.79 and 272 pg/ml. AUC of presepsin was larger than AUCs of WBC, CRP and NLR, suggesting the highest predicting power of presepsin among other parameters.Conclusion: Plasma presepsin level with CRP, WBC and NLR are valuable diagnostic parameters to predict complicated acute appendicitis.

Determine White Blood Cell Count (WBCC) Limits and ABL Ct Cut-Off with ABL Copy Number (CN) Calibration for the Xpert® BCR-ABL Ultra Assay

Introduction Monitoring BCR-ABL transcript levels in peripheral blood of patients using real-time quantitative PCR is standard of care in the management of Chronic Myeloid Leukemia. Xpert BCR-ABL Ultra is an automated cartridge-based assay developed by Cepheid for sensitive assessment of BCR-ABL transcript levels. However, in clinical situations the total RNA isolated from white blood cell (WBC) in the patient's blood is insufficient to ensure assay LoD of 0.003% (IS)/MR4.52, or high WBCC in specimen can overload the Ultra cartridge, resulting in inaccurate test results. Therefore, it is necessary to determine proper WBCC input, to ensure accurate assay quantification results. Objectives The objective of this study was to determine the correlation between ABL copy number input and Ct value, determine lower and upper WBCC input limits for the Xpert BCR-ABL Ultra assay and establish the ABL Ct cut-offs. Methods To establish an upper ABL Ct limit that corresponds to a minimal ABL CN input of 32,000 for supporting LoD, a BCR-ABL plasmid DNA (pDNA, ERM-AD623) was serially diluted and tested in an EDTA whole blood background to make a calibration panel. A standard curve was generated against which the ABL Ct could be estimated as a function of ABL copy number. To estimate the lower WBCC limit, CML negative EDTA lysate with normal WBCC was serially diluted with WBC depleted EDTA lysate. To estimate the upper WBCC limit, EDTA lysate from a specimen with high WBCC (69.9×106cells/mL) was diluted with CML negative lysate with normal WBCC (3.5×106 cells/mL). To validate the WBCC limit, CML positive lysate with high WBCC (40×106cells/mL) was diluted with high WBCC (40×106cells/mL) CML negative lysate to produce five BCR-ABL levels within 0.03% (IS)/MR3.52 to 65% (IS)/MR0.19. Each level was further diluted with PBS lysate to achieve 8 levels of WBCC from 40×106 to 0.04×106 cells/mL. Results ABL Ct value of ~19 corresponding to a minimum ABL copy number input of 32,000 was established to support assay LoD of 0.003% (IS)/MR4.52 (Figure.1). To allow for a one Ct buffer, the upper ABL Ct cut-off was set at 18. The lower WBCC input limit, corresponding to the upper ABL Ct limit of ~18 with estimated ABL copy number greater than 64,000, was established to be 0.15×106 cells/mL (Figure.2) The upper WBCC input limit was established to be 30×106 cells/mL. This corresponds approximately to a Ct value of 10 (Figure.3). The Xpert BCR-ABL Ultra test monitors RNA expression levels of the BCR-ABL fusion transcript and the ABL transcript. Unlike a gene DNA marker, RNA expression levels are variable and can fluctuate. To accommodate for the variability in RNA expression levels and based on results from this study, the upper ABL Ct cut-off was lowered to 8. The lower and upper WBCC input limit, together with ABL Ct cut-off, were validated with CML patient specimens with WBCC input ranging from 0.04 to 40 × 106 /ml within IS% range of 0.03% to 65% (Table.1). The normal WBCC of 4 to 10 × 106/mL across all tested concentrations yielded ABL Ct range from 13.2 to 11.7 with estimated ABL copy number of 2.5×106 to 7.8×106. Conclusions A correlation between ABL copy number input and ABL Ct value was established with the standard curve generated from a pDNA calibration panel. The lower WBCC limit was set at 1.5×105 cells/mL and upper WBCC limit was set at 3.0×107 cells/mL, together with ABL Ct cut-offs of 18 and 8 and ABL copy number of 6.4×104 to 1.4×108, to support Xpert BCR-ABL Ultra assay performance. The normal WBCC of 4 to10×106/mL across all tested concentrations yielded an ABL Ct range from 13.2 to 11.7 with estimated ABL copy number of 2.5×106 to 7.8×106, indicating Xpert BCR-ABL Ultra assay normally provided more than 10 times the required minimum ABL Copy number to support the assay LoD claim at 0.003% (IS). *Product in development. Not for use in diagnostic procedures. Not reviewed by any regulatory body. Disclosures No relevant conflicts of interest to declare.

Right ventricular failure following left ventricular assist device implantation is associated with a preoperative pro-inflammatory response

BackgroundSystemic inflammation during implant of a durable left ventricular assist device (LVAD) may contribute to adverse outcomes. We investigated the association of the preoperative inflammatory markers with subsequent right ventricular failure (RVF).Materials and methodsProspective data was collected on 489 patients from 2003 through 2017 who underwent implantation of a durable LVAD. Uni- and multivariable correlation with leukocytosis was determined using linear and binary logistic regression. The population was also separated into low (< 10.5 K/ul, n = 362) and high (> 10.5 K/ul, n = 127) white blood cell count (WBC) groups. Mantel-Cox statistics was used to analyze survival data.ResultsPostop RVF was associated with a higher preop WBC (11.3 + 5.7 vs 8.7 + 3.1) and C-reactive protein (CRP, 5.6 + 4.4 vs 3.3 + 4.7) levels. Multivariable analysis identified an independent association between increased WBC preoperatively with increased lactate dehydrogenase (LDH, P < 0.001), heart rate (P < 0.001), CRP (P = 0.006), creatinine (P = 0.048), and INR (P = 0.049). The high WBC group was more likely to be on preoperative temporary circulatory support (17.3% vs 6.4%, P < 0.001) with a trend towards greater use of an intra-aortic balloon pump (55.9% vs 47.2%, P = 0.093). The high WBC group had poorer mid-term survival (P = 0.042).ConclusionsPostop RVF is associated with a preoperative pro-inflammatory environment. This may be secondary to the increased systemic stress of decompensated heart failure. Systemic inflammation in the decompensated heart failure may contribute to RVF after LVAD implant.

The Role of Regulatory B Cells in Patients with Acute Myeloid Leukemia.

BackgroundRegulatory B (Breg) cells are a group of B cells with immunomodulatory function, which mainly exert negative immunomodulatory function by secreting IL-10 and other cytokines. Due to their immunoregulatory properties, Breg cells may participate in the pathogenesis of acute myeloid leukemia (AML). This study was designed to explore the frequency of Breg cells and the relationship between the Breg cells and clinical data in patients with AML.Material/MethodsA total of 46 (36 in peripheral blood, 10 in bone marrow) AML patients and 15 healthy donors (HD) were included for detection of Breg cells frequency by multicolor flow cytometry. All cases were divided into different groups according to FAB subtypes of leukemia, white blood cell count (WBC) levels, age, cytogenetic characteristics, and molecular abnormalities, and were compared the differences of Breg cell frequency. Survival curve analysis was performed to estimate the value of Breg cell frequency in prognosis among cases with AML.ResultsWe found that the frequency of Breg cells was higher in AML patients both in peripheral blood (PB) and in bone marrow (BM) compared with those in HDs. The AML patients with high WBC levels had higher Breg cell frequency compared with those with low WBC levels. Low-risk patients with had lower Breg cells frequency compared to the medium-risk patients. The patients with high WBC and high Breg cells frequency showed a shorter overall survival. Similarly, the overall survival of AML patients in the older group with high Breg cells frequency was significantly shorter than in the younger group with low Breg cell frequency.ConclusionsFor AML patients, the frequency of Breg cells was elevated, and high frequency of Breg cells may reveal poor prognosis.

White Blood Cell Count and C-Reactive Protein Independently Predicted Incident Diabetes: Yuport Medical Checkup Center Study

ABSTRACTAim: White blood cell (WBC) count or C-reactive protein (CRP) level alone may not fully indicate the chronic inflammation causing type 2 diabetes. We examined both WBC count and CRP level, independently and in combination, as predictive markers for type 2 diabetes and also considered the influence of obesity and other individual characteristics on the relationship.Materials and Methods: In total, 9,706 participants were enrolled with WBC < 10*109/L and CRP < 10 mg/L using data from the Yuport Medical Checkup Center Study. The cumulative incidence of type 2 diabetes [defined either as known diabetes, fasting plasma glucose ≥ 7.0 mmol/L, or HbA1c ≥ 6.5% (47.5 mmol/mol)] was measured. Hazard ratios (HRs) were estimated using a Cox proportional hazards model.Results: During study period, 272 men (5.5%) and 113 women (2.4%) progressed to diabetes. The progression to diabetes was predicted by both increased baseline levels of WBC count [adjusted HR = 1.29 (95% CI: 1.04–1.60)] and CRP level [1.39 (1.10–1.74)], even after adjusting for possible confounders. The combined presence was more predictive of diabetes than either alone in a four-groups analysis [1.75 (1.28–2.40)]. In addition, the elevated HRs of either or both higher WBC and CRP levels were observed across four subgroups of body mass index (BMI), including low BMI, and people who had at least one occurrence of dyslipidemia.Conclusion: Increased WBC counts and CRP levels were predictive for type 2 diabetes and the combination augmented the risk of diabetes, regardless of whether the BMI was high or low.

Combined utility of white blood cell count and blood glucose for predicting in-hospital outcomes in acute ischemic stroke

BackgroundHigh white blood cell (WBC) count and high blood glucose level are risk factors for mortality and pneumonia after acute ischemic stroke (AIS). We investigated the combined effect of high WBC count and high blood glucose level on hospital admission and in-hospital mortality and pneumonia in acute AIS patients.MethodsA total of 3124 AIS patients enrolled from December 2013 to May 2014 across 22 hospitals in Suzhou city were included in the present study. We divided patients into four groups according to their level of WBC count and blood glucose: NWNG (normal WBC count and normal glucose), NWHG (normal WBC count and higher glucose), HWNG (higher WBC count and normal glucose), and HWHG (higher WBC count and higher glucose). Cox proportional hazard model and logistic regression model were used to estimate the combined effect of WBC count and blood glucose on all-cause in-hospital mortality and pneumonia in AIS patients.ResultsHWHG was associated with a 2.22-fold increase in the risk of in-hospital mortality in comparison to NWNG (adjusted hazard ratio [HR] 2.22; 95% confidence interval [CI], 1.21–4.07; P trend = 0.003). The risk of pneumonia was significantly higher in patients with HWHG compared to those with NWNG (adjusted odds ratio [OR] 2.61; 95% CI, 1.66–4.10; P trend < 0.001). The C-statistic for the combined WBC count and blood glucose was higher than WBC count or blood glucose alone for prediction of in-hospital mortality and pneumonia (all p < 0.01).ConclusionsHigh WBC count combined with high blood glucose level at admission was independently associated with in-hospital mortality and pneumonia in AIS patients. Moreover, the combination of WBC count and blood glucose level appeared to be a better predictor than WBC count or blood glucose alone.