Nucleated Red Blood Cell Counts Research Articles

Validation of the Sysmex XN-V hematology analyzer for feline specimens.

The Sysmex XN-V is derived from the new Sysmex XN series of human hematology analyzers. The main changes from the previously validated XT-2000iV analyzer include an optic-fluorescent analysis for platelets and a nucleated red blood cell (NRBC) count. We aimed to validate the Sysmex XN-V for feline blood following the American College for Veterinary Clinical Pathology and International Council for Standardization in Hematology recommendations. Feline EDTA blood specimens were analyzed on the Sysmex XN-V to evaluate repeatability, linearity, comparison with the XT-2000iV analyzer and manual methods, stability, and to verify the previously established Sysmex XT-2000iV RIs. Repeatability was excellent for most variables. Visually determined linearity was excellent or good for most variables except eosinophils and platelet variables. The correlation between the XN-V and XT-2000iV analyzers was good (≥0.82) for all variables except reticulocyte indices. Correlations between the Sysmex XN-V and manual differential counts were good to excellent for most variables, acceptable for neutrophils, and fair for monocytes and NRBC. The previously established Sysmex XT-2000iV RIs can be used to interpret results from the Sysmex XN-V analyzer for most variables except red cell distribution width and reticulocyte variables. The RI for platelet variables could not be evaluated because of platelet clumps. Changes in the Sysmex XN-V measurements after storage at 4 and 24°C were similar to those described for the Sysmex XT-2000iV analyzer. The performance of the Sysmex XN-V analyzer was good and compared favorably with the Sysmex XT-2000iV analyzer.

Comparative assessment of nucleated red blood cells using a fully automated hematology analyzer versus slide examination.

Objective: To determine the sensitivity and specificity of fully automated hematology analyzer for nucleated red blood cells (NRBCs) keeping microscopic slide examination as gold standard. Study Design: Cross-sectional study. Setting: Department of Hematology, RMI Hospital, Peshawar. Period: 1st December 2023 to 29th Feb 2024. Methods: A total of 160 blood samples referred for assessment to the hematological department for the suspected abnormal NRBCs were included in the study through consecutive sampling. Fresh Blood samples were analyzed using fully automated hematology analyzer. Manual counting of the NRBCs of the same samples was done using manual blood smear slide examination. Findings of the automated nucleated red blood cell counting methods were statistically analyzed keeping microscopic slide examination as gold standard. Sensitivity, specificity, accuracy and negative and positive predictive values were calculated for diagnosis of NRBC (count >0.02 × 109/L). Clinical sensitivity of the test was determined by Receiver Operating Test (ROC). Results: The Mean±SD of patient’s age in this study was 46.78±14.89 years with an age range of 0.5 to 67 years. The male gender was 53.75% while female gender was 46.25%. Fully automated hematology analyzer detected 30 (18.75%) samples while slide examination detected 31 (19.37%) samples with NRBC count >0.02 × 109/L. Fully automated hematology analyzer has shown sensitivity of 93.55 %, specificity 99.23% and diagnostic accuracy by 98.13%. PPV was 96.67% and NPV was 98.46 (p < 0.000) for the diagnosis of NRBC. The cutoff value of 0.0175 × 109/L (20/µL) offered the best balance between sensitivity and specificity based on ROC curve. Conclusion: Association is present for the NRBC count among the fully automated hematology analyzer and microscopic slide examination.

Comparison Evaluation of Automated Nucleated Red Blood Cell Enumeration by Sysmex XN 1000 in Comparison With Microscopic Reference in Children Under 1 Year.

In newborns, elevated nucleated red blood cell (NRBC) levels can be associated with enhanced erythropoietic stress and might be predictive for adverse outcome. Also, the presence of NRBC in peripheral blood might lead to erroneous enumeration results of white blood cells in automated hematology analyzers. We aimed to assess the comparability of the Sysmex XN 1000 to manual slide reviews and correlation of NRBC with inflammation markers. Specimens of 3397 children under 1 year were compared by automated and microscopic NRBC enumeration. Additionally, potential correlations between NRBC and age and inflammation markers were examined. Overall, there was good correlation (r = 0.97) between automated (range: 0%-3883%) and microscopic enumeration (range: 0%-3694%) of NRBC with high comparability up to a NRBC value of 200% and an increase in the variation between the two methods with increasing NRBC numbers. When 94 samples with ≤ 200% NRBC and ≥ 30% divergence between methods were separately reanalyzed with respect to overlapping cell populations in their scattergrams, Sysmex would have generated unrecognized incorrect automated results in 47 samples, corresponding to 1.4% of total study samples. NRBC counts were negatively correlated to age, but not to inflammation markers. Sysmex XN 1000 is highly precise in the enumeration of NRBC in children under 1 year up to counts of 200% and might replace time-intense manual counting in routine diagnostics. In the setting of neonatal and intensive care diagnostics, microscopic control and supervision of scattergrams are highly recommended for any automated NRBC enumeration processes.

Correlation of nucleated red blood cell counts by Sysmex XN-1000 with conventional microscopy

Objective: To correlate the results of manual microscopy with that of Sysmex XN-1000 analyzers and to determine its diagnostic accuracy, taking microscopy as gold standard. Material and Methods: The study was conducted at Chughtai Institute of Pathology from July 1st -December 31st, 2022. Nucleated Red Blood Cells (nRBCs) were manually enumerated against 100 White Blood Cells (WBCs) on peripheral smears and 511 such positive cases were included. 500 samples negative for nRBCs by manual microscopy were also included. These positive and negative samples were run on two Sysmex XN-1000 analyzers. A result of 0.3 and above nRBCs/100 WBCs on Sysmex XN-1000 was considered positive and less than 0.3 as negative. Mean values of both manual and automated methods were correlated. Results: Our study mentioned nRBCs ranging from 0 to 1330 nRBCs/100WBCs by manual count. Compared with Sysmex XN-1000 analysis, the Deming regression analysis showed the mean difference between the two methods was 0.055 (95%CI: −0.015 to +0.124) (p>0.05 no proportional bias between two methods). The Sysmex XN produced an area under the curve, by ROC analysis, at 0.894 (P < .001). The overall sensitivity was (90.56%) (CI 87.56-92.98%), with few false-negative results. The overall specificity is (88.30%) (CI 85.20%-90.96%) due to few false positivity. Positive predictive value and negative predictive value were 88.26% (CI 88.54-90.52%) and 90.6% (CI 87.99-92.69%) respectively. Accuracy was 89.42% (CI87.35-91.25%). Conclusion: Sysmex XN1000 analyzer is a highly sensitive and accurate instrument which can replace nRBCs enumeration using manual counting by conventional microscopy. Keywords: Nucleated red blood cells, Hematology analyzer, Sysmex XN 1000, Manual microscopy

Comparison of atlantoaxial and lumbosacral cerebrospinal fluid centesis techniques in South American camelids.

Iatrogenic blood contamination during cerebrospinal fluid (CSF) centesis is common, which can limit the diagnostic usefulness of the sample. A novel ultrasound-guided CSF collection technique is described in horses, by which CSF is obtained from the atlantoaxial (AA) space. To compare ultrasound-guided AA centesis with lumbosacral (LS) centesis in South American camelids (SAC). The hypotheses were that AA centesis would yield samples with less blood contamination although being technically more challenging than LS centesis. Eight clinically healthy adult SAC from a university-owned teaching herd. Single-blinded, randomized, 4-way, 4-period crossover study in which 2 veterinarians each performed both centesis techniques on each animal once. Cytological sample analysis was performed, and the technical difficulty of sample acquisition was assessed. The CSF was collected successfully and without complications by either technique during all collection attempts. Aspects of technical difficulty and concentrations of CSF analytes did not vary significantly between techniques. Median total nucleated cell and red blood cell counts were 1/μL and 0.5/μL and 167.5/μL and 155/μL for AA and LS techniques, respectively. The median total protein concentration was 32.9 mg/dL and 38 mg/dL for AA and LS centeses. A median of 1 attempt was necessary for both centesis techniques and the median number of needle repositioning events was 1 for AA and 0 for LS. Depending on clinical circumstances, ultrasound-guided AA centesis appears to be an acceptable alternative to other techniques for collection of CSF from SAC.

Nucleated Red Blood Cell Counts Differentiate Cardiac from Respiratory Causes of Cyanosis at Birth.

Tissue hypoxia increases erythropoietin production and release of immature erythrocytes that can be measured using nucleated red blood cell counts (nRBC). We hypothesized that hypoxia due to congenital heart disease (CHD) is chronic and is better tolerated than hypoxia due to respiratory disease (RD), which is an acute stress in newborns leading to higher nRBC. This study assesses the utility of nRBC as a marker to differentiate hypoxia due to CHD vs RD in term neonates. This was a single-center, retrospective study of term neonates with cyanosis from 2015 to 2022. Neonates < 37weeks of gestation, with hypoxic-ischemic encephalopathy, and those with other causes of cyanosis were excluded. The patients were divided into 2 groups: cyanotic CHD and cyanotic RD. Clinical and laboratory data done within 12h and 24-36h after birth were collected. Data are represented as median and Interquartile range. Of 189 patients with cyanosis, 80 had CHD and 109 had RD. The absolute nRBC count at ≤ 12h of age was lower in the CHD (360 cells/mm3) compared to RD group (2340 cells/mm3) despite the CHD group having significantly lower baseline saturations. A value of 1070 cells/mm3 was highly sensitive and specific for differentiating CHD from RD. The positive predictive value for this cut-off value of 1070 cells/mm3 was 0.94 and the negative predictive value was 0.89. The absolute nRBC is a simple screening test and is available worldwide. A nRBC < 1070 cells/mm3 in cyanotic newborns should hasten the search for CHD etiology with the possible need for prostaglandin therapy.

Hematological profiles of newborns of mothers with hypertensive disorders of pregnancy delivered at the University of Gondar comprehensive specialized hospital: a comparative cross-sectional study

BackgroundHypertensive disorders in pregnancy can cause prenatal placental perfusion with insufficient blood supply to the fetus, resulting in fetal exposure to hypoxia and leading to disturbance of neonatal hematopoietic stem cells. This study aimed to compare the hematological profiles of newborns from mothers with hypertensive disorders and normotensive delivered at the University of Gondar comprehensive specialized hospital.MethodsA comparative cross-sectional study was conducted from March to May 2022 among 308 newborns from hypertensive and normotensive mothers in equal proportions. A systematic random sampling technique was used to select study participants. Three milliliters of cord blood were collected to perform a complete blood count by Beckman coulter. The results were presented using tables and graphs. Independent t-test and Mann-Whitney U test were done to compare the hematological profiles of the two groups. P-value < 0.05were considered statistically significant.ResultsThe majority of hypertensive and normotensive mothers’ ages were between 20 and 34 years (83.77% and 90.91%, respectively). The hematocrit levels were significantly higher in neonates of hypertensive mothers than the neonates of normotensive mothers (49.10 ± 5.19% and 46.09 ± 7.63% respectively) (P < 0.001) while neutrophil counts were significantly lower in neonates of hypertensive mothers than the neonates of normotensive mothers (6.62 ± 3.30 and 7.55 ± 3.31 × 103 /ul respectively) (P = 0.007). Also, platelets counts were significantly lower in neonates of hypertensive mothers than neonates of normotensive mothers (221.25 ± 83.56 and 260.24 ± 83.01 × 103/ul respectively) (P < 0.001). The platelet and nucleated red blood cell count showed a statistically significant difference among newborns from mothers with superimposed preeclampsia and gestational hypertension.ConclusionNewborns delivered from hypertensive disorders of pregnancy had low white blood cell parameters, low platelet count and high red blood cell parameters compared to controls. As result, newborns may develop leukopenia, thrombocytopenia and polycythemia, respectively. Therefore, newborns should be monitored for early detection and follow-up of hematological abnormalities before complications occurred.

Complete Blood Count Reporting Among Hospitals in the United States

Background Complete blood count (CBC) is the most common test performed in hematology-oncology. A typical CBC report contains over 10 elements. Simplifying the report by removing nonessential elements may benefit clinicians and patients. Little is known about what elements are reported by various hospitals in the US. The objective of our study was to describe how hospitals in the US report CBC. Methods After approval from the Mayo Clinic institutional review board, we obtained hospital CBC reported elements from the Epic electronic medical records Care Everywhere health information exchange. CBCs from 2020-2023 were included. For each hospital, we determined the city and state where it's located. We obtained the number and type of elements reported in the CBC and compared the findings between academic and community hospitals. Results We collected CBC data from 139 hospitals across 43 states and 102 cities (Figure 1). Fifty-four percent were community hospitals while 46% were academic hospitals. The median number of elements reported by all hospitals was 21 (range, 12-24). This was not different (P=0.98) between academic (median 22 [range, 12-24]) and community hospitals (median 21 [range, 14-24]; Figure 2). The distribution of hospitals according to the number of CBC elements reported were as follows: 12-15 (6%), 16-19 (13%), 20 (25%), 21(6%), 22 (18%), 23 (19%), and 24 (13%). A minority of the hospitals did not report % white blood cell differential count (9.4%) or mean platelet volume (20.9%). Nucleated red blood cell and immature granulocyte count were reported frequently, 46.0% and 56.8%, respectively. However, the frequency of reporting these four elements were similar between academic and community hospitals. Conclusions We found substantial variations in CBC reporting among US hospitals even within academic and community hospitals. Most hospitals reported 20 or more elements, twice as many as the hospitals reporting the least number of elements. There is an opportunity to simplify and standardize CBC reporting, which will minimize physician burnout and distraction as well as decrease patient concern and query.

Identification, Monitoring and Reversibility of PARP Inhibitor Associated Clonal Haematopoiesis and Myelodysplastic Syndrome: A Case Series

Background: Poly ADP ribose polymerase inhibitors (PARPi) are widely used as maintenance treatment for ovarian cancer (OC) after response to platinum chemotherapy in the first line setting and recurrent disease. Their use has significantly improved outcomes, particularly in patients with BRCA mutations in whom first-line PARPi maintenance therapy increases progression-free survival from 13.8 to 56.0 months (Hazard Ratio, 0.33; 95% CI, 0.25-0.43). However, a recent meta-analysis of clinical trial adverse events and pharmacovigilance data showed that PARPi therapy significantly increases the risk of therapy related myeloid neoplasms (tMN), myelodysplastic syndrome (MDS) and acute myeloid leukaemia with poor outcomes. Platinum based chemotherapy is the mainstay of OC treatment and is associated with clonal haematopoiesis (CH) with mutations in DNA damage response genes. CH is recognised as a premalignant myeloid state but real world data on clonal dynamics during PARPi therapy are lacking. We present a case series of OC patients receiving PARPi-therapy referred for haematological review for beyond-expected HT or unexplained full blood count (FBC) abnormalities with serial FBC and myeloid next generation sequencing (mNGS) clonal monitoring in relation to PARPi therapy. Methods: Seven patients with stage III/IV high-grade OC were referred to haematology at the discretion of the treating oncologist between February 2017 and May 2022 (mean age (SD): 68 (7.0) years, median PARPi duration: 18 months (range: 4-86)). All patients had received prior platinum-based chemotherapy and were screened with mNGS in peripheral blood (PB) or bone marrow aspirate and trephine (BMAT) at initial assessment. Results: Reasons for referral are detailed in Table 1 and include two patients with isolated elevated nucleated red blood cell (NRBC) counts. Six patients had clonal pathology (CP), five of whom had single-nucleotide variants typical of platinum-exposure defined by the involved gene or platinum-associated mutational signatures. Clinical details including BRCA-status, haematological diagnosis and mNGS results are detailed in Table 1. PARPi-therapy was discontinued in four patients with CP, with resolution or improvement of FBC and m-NGS abnormalities (variant allele frequencies (VAFs) and/or copy number variations (CNVs)) in three patients. These findings were most marked in Patient 1, referred with grade 3 thrombocytopenia and transfusion dependence and in whom PARPi cessation resulted in transfusion independence and resolution of morphological and cytogenetic abnormalities (clinical course including BMAT results and haematological parameters illustrated in Figure 1), and Patient 3, who demonstrated selective regression of a TP53 mutated clone on PARPi cessation. Two patients with germline BRCA mutations continued PARPi therapy with 3 monthly PB mNGS monitoring after discussion at multidisciplinary team meetings. Conclusions: We show using sequential molecular and haematological monitoring before and after PARPi cessation that morphological, FBC and clonal abnormalities can be reversed on PARPi discontinuation, potentially mitigating the risk of life-threatening tMNs in some patients. We also highlight that, in addition to cytopenias, persistently elevated NRBCs may be an indicator of clinically relevant CP not well described in pharmacovigilance data based on common terminology criteria for adverse events criteria. Large scale prospective data on clonal dynamics in PARPi therapy are needed to inform mNGS screening and tMN risk stratification as scoring systems based on age associated clonal haematopoiesis of indeterminate potential may not be applicable. Until these data are available we demonstrate the value in MDT discussion and serial PB mNGS monitoring in formulating case-by-case therapeutic decisions regarding CP and PARPi therapy, particularly patients with high risk BRCA mutated OC.

Nucleated red blood cells as a novel biomarker in the diagnosis and prediction of sepsis severity in children.

Sepsis is a vitally serious disease leading to high mortality. Nucleated red blood cells (NRBCs) are present in some noninfectious diseases, but the relationship between NRBCs and sepsis in children remains unknown. The purpose of this study was to compare the clinical characteristics and outcomes of sepsis with positive NRBCs and negative NRBCs in children, and to further explore whether the count of NRBCs has a relationship with the severity of sepsis. We enrolled children with sepsis who were admitted to the Children's Hospital of Chongqing Medical University between January 2020 and December 2022. The children's clinical data, laboratory data and outcomes were recorded and analyzed. One hundred and fifteen children met the inclusion criteria in our study. Compared to negative NRBCs patients, the C-reactive protein, alanine transaminase, urea nitrogen values, mortality rate and length of hospitalization were found to be significantly increased, while platelet counts, and hemoglobin were significantly decreased in sepsis patients with positive NRBC (P < 0.05). Receiver operating characteristic (ROC) curves analysis showed that the optimal cutoff value of the NRBC count in the diagnosis of severe sepsis was 3, with a sensitivity of 87.5% and specificity of 94.9%. The area under the ROC curve was 0.877 (95% CI: 0.798-0.957). These findings demonstrated that NRBC count has the potential to be a biomarker for the diagnosis of sepsis in children, especially an NRBC count greater than 3, which may predict the severity and poor prognosis in children suffering from sepsis.

Nucleated Red Blood Cell Emergence-Time in Newborn Lambs Following a Dose of Darbepoetin Alfa.

Nucleated red blood cells (NRBC) are very uncommon in the blood of children and adults, but small numbers are not rare in neonates on the day of birth. Elevated NRBC counts in neonates can be seen following erythropoietin dosing. Limited studies in human neonates suggest the time-interval between erythropoietin dosing and the first appearance of NRBC in the blood (the "NRBC emergence-time") is in excess of 24 hours. We made serial blood counts (Sysmex veterinary analyzer) on ten newborn lambs; five were dosed with darbepoetin (10 µg/kg), and five were dosed with a vehicle-control to assess the NRBC emergence time under relatively controlled laboratory conditions. The first appearance of NRBC was at 24 h (2757±3210 NRBC/µL vs. 0/µL in controls). Peak was 48-72 h (16,758±8434/µL vs. 0/µL in controls), followed by fewer at 96 hours (7823±7114/µL vs. 0/µL in controls). Similarly, reticulocytes peaked at 48-72 h (113,094±3210/µL vs. 10,790±5449/µL in controls), with no changes in platelets or leukocytes. The NRBC emergence time in newborn lambs is similar to reports from newborn humans. By extrapolation, if a neonate has a high NRBC at birth, the erythropoietic stimulus likely occurred within the interval 24 to perhaps 96+ hours prior to birth.

Nucleated red blood cells in neonates with hypoxic ischaemic encephalopathy treated with hypothermia: A worthwhile prognostic biomarker for clinicians in LMIC?

Background. Neonatal hypoxic ischaemic encephalopathy (HIE) is a leading cause of term neonatal death worldwide, with a higherincidence in low- to middle-income settings.Objective. To investigate whether nucleated red blood cell (nRBC) counts could predict severity of HIE and outcomes in term neonatestreated with therapeutic hypothermia (TH).Methods. We conducted a retrospective sub-study at Tygerberg Hospital in Cape Town, South Africa. The review included all cooledneonates’ clinical records and blood samples from a National Health Laboratory Services database. One experienced neurodevelopmental expert assessed patients over a period of 12 months.Results. Twenty-five files out of a total of 100 were excluded owing to missing data. In accordance with the Thompson HIE score,the cohort was classified as mild (56%), moderate (27%), and severe (17%). All included patients (n=75) had full blood counts within6 hours of delivery. nRBC were detected in 52% of the samples. There was no correlation between nRBC category and HIE severity(p=0.265). Raised nRBCs (≥30 cells/100 white blood cells (WBCs)) were more frequent in infants who died than in those whosurvived (p=0.008). Infants with nRBC counts ≥30 cells/100 WBCs had an increased likelihood of having cerebral palsy or impairedneurodevelopment (p=0.013).Conclusion. The study demonstrated a significant association between an early increase in nRBC counts in HIE infants treated with TH,and both short- and long-term outcomes. A larger multicentre study is required to better understand the relationship between nRBCcounts and HIE in the era of cooling in our local setting.

Machine learning of cell population data, complete blood count, and differential count parameters for early prediction of bacteremia among adult patients with suspected bacterial infections and blood culture sampling in emergency departments.

Bacteremia is a life-threatening complication of infectious diseases. Bacteremia can be predicted using machine learning (ML) models, but these models have not utilized cell population data (CPD). The derivation cohort from emergency department (ED) of China Medical University Hospital (CMUH) was used to develop the model and was prospectively validated in the same hospital. External validation was performed using cohorts from ED of Wei-Gong Memorial Hospital (WMH) and Tainan Municipal An-Nan Hospital (ANH). Adult patients who underwent complete blood count (CBC), differential count (DC), and blood culture tests were enrolled in the present study. The ML model was developed using CBC, DC, and CPD to predict bacteremia from positive blood cultures obtained within 4h before or after the acquisition of CBC/DC blood samples. This study included 20,636 patients from CMUH, 664 from WMH, and 1622 patients from ANH. Another 3143 patients were included in the prospective validation cohort of CMUH. The CatBoost model achieved an area under the receiver operating characteristic curve of 0.844 in the derivation cross-validation, 0.812 in the prospective validation, 0.844 in the WMH external validation, and 0.847 in the ANH external validation. The most valuable predictors of bacteremia in the CatBoost model were the mean conductivity of lymphocytes, nucleated red blood cell count, mean conductivity of monocytes, and neutrophil-to-lymphocyte ratio. ML model that incorporated CBC, DC, and CPD showed excellent performance in predicting bacteremia among adult patients with suspected bacterial infections and blood culture sampling in emergency departments.

Diagnostic Value and Prognostic Significance of Nucleated Red Blood Cells (NRBCs) in Selected Medical Conditions.

Nucleated red blood cells (NRBCs) are premature erythrocyte precursors that reside in the bone marrow of humans of all ages as an element of erythropoiesis. They rarely present in healthy adults' circulatory systems but can be found circulating in fetuses and neonates. An NRBC count is a cost-effective laboratory test that is currently rarely used in everyday clinical practice; it is mostly used in the diagnosis of hematological diseases/disorders relating to erythropoiesis, anemia, or hemolysis. However, according to several studies, it may be used as a biomarker in the diagnosis and clinical outcome prognosis of preterm infants or severely ill adult patients. This would allow for a quick diagnosis of life-threatening conditions and the prediction of a possible change in a patient's condition, especially in relation to patients in the intensive care unit. In this review, we sought to summarize the possible use of NRBCs as a prognostic marker in various disease entities. Research into the evaluation of the NRBCs in the pediatric population most often concerns neonatal hypoxia, the occurrence and consequences of asphyxia, and overall neonatal mortality. Among adults, NRBCs can be used to predict changes in clinical condition and mortality in critically ill patients, including those with sepsis, trauma, ARDS, acute pancreatitis, or severe cardiovascular disease.

Analytical errors in nucleated red blood cell enumeration.

Enumeration of nucleated red blood cells (NRBCs) in peripheral blood of dogs and cats is performed by manual counting during blood film evaluation. Automated methods have increased precision and accuracy; however, most analyzers cannot distinguish leukocytes and NRBCs. The Sysmex XN-V Series may distinguish NRBCs and leukocytes; however, analytical errors occur. We aimed to investigate cases with discrepant automated and manual NRBC counts, and to evaluate reasons for analytical errors. Data from samples with increased NRBCs were collected retrospectively and compared with manual counts performed on blood films using Spearman's correlation, Passing-Bablok agreement analysis, and Bland-Altman comparisons. Precision of the automated method and interobserver agreement of manual counts were evaluated. Cases with discrepant results were investigated. Agreement between the methods was good at ≤1NRBC ×109 /L in dogs and cats, and inadequate at ≥1NRBC ×109 /L. The automated method demonstrated a negative proportional difference to the manual method. Precision was good for the automated method (overall CV 7.1%) and interobserver agreement for the manual method was poor overall (mean CV 27.3%, range 0%-106.1%). Inaccuracies in NRBC enumeration by the automated method occurred with high hematocrits, the mergence of the cell fragments and leukocyte clouds, and the presence of earlier erythroid precursors. NRBC enumeration by the WNR channel on the Sysmex XN-1000 V is precise and has good agreement with manual counts in canine and feline blood samples at ≤1NRBC ×109 /L. Manual film review is indicated for samples with ≥1NRBC ×109 /L, earlier erythroid precursors, samples from greyhounds and dehydrated patients, and if gating errors are noted.

Comprehensive pediatric reference intervals for 79 hematology markers in the CALIPER cohort of healthy children and adolescents using the Mindray BC-6800Plus system.

Hematological parameters vary significantly throughout growth and development due to physiological processes such as fetal-to-adult erythropoiesis and puberty. Pediatric age- and sex-specific reference intervals (RIs) are thus essential for appropriate clinical decision-making. The current study aimed to establish RIs for both common and novel hematology parameters on the Mindray BC-6800Plus system. Six hundred and eighty-seven healthy children and adolescents (30 days to 18 years) were enrolled. Participants were recruited as part of the Canadian Laboratory Initiative on Pediatric Reference Intervals Program upon informed consent or identified from apparently healthy outpatient clinics. Whole blood was collected and assayed for 79 hematology parameters on the BC-6800Plus system (Mindray). Age- and sex-specific RIs were established as per Clinical and Laboratory Standards Institute EP28-A3c guidelines. Dynamic reference value distributions were observed for several hematology parameters, including erythrocytes, leukocytes, platelets, reticulocytes, and research-use-only markers. Age partitioning was required for 52 parameters, demonstrating changes in infancy and puberty. Sex partitioning was required for 11 erythrocyte parameters (i.e., red blood cell (RBC), hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin concentration, RBC distribution width coefficient of variation, hemoglobin distribution width, macrocyte count, macrocyte percentage, RBC (optical), and reticulocyte production index). Few parameters had undetectable levels in our healthy cohort (i.e., nucleated RBC count and immature granulocyte count). The current study completed hematological profiling for 79 parameters on the BC-6800Plus system in a healthy cohort of Canadian children and adolescents. These data emphasize the complex biological patterns of hematology parameters in childhood, particularly at the onset of puberty, and support the need for age- and sex-specific RIs for clinical interpretation.

Nucleated red blood cell: a feasible quality parameter of cord blood units

IntroductionUmbilical cord blood is an alternative source of hematopoietic progenitor cells for bone marrow transplantation; however, it is associated with a higher graft failure rate. The presence of a high rate of nucleated red blood cells (NRBCs) seems to be related to a greater capacity for engraftment, although is also associated with fetal distress conditions. We analyzed the correlation of the NRBC with quality parameters and its association with the utilization score of a cord blood unit. Study design and methodData of 3346 units collected in a public cord blood bank from May 2010 to December 2017 were analyzed, retrospectively, to identify factors associated with an increased number of nucleated red blood cells and its correlation with the engraftment capacity measured through total nucleated cells (TNCs) and CD34 positive cells. We also evaluated the utilization score of these units and identified an NRBC cutoff associated with a higher score. ResultsThe median volume collected was 104 mL (42–255), the pre-processing TNC count was 144.77 × 107 (95.46–477.18), the post-processing TNC count was 119.44 × 107 (42.7–477.18), the CD34 count was 4.67 × 106 (0.31–48.01), the NRBC count was 5 (0–202) and the utilization score was 0.0228 (0.00143–0.9740). The NRBC showed a correlation with the collected volume, TNC and CD34 positive cells and a higher utilization score and the receiver operating characteristic (ROC) curve analysis identified the five NRBC/100 leukocytes cutoff that correlates better with the probability of use. No association with pathological conditions and the NRBC rate was observed. ConclusionsThe NRBC is a feasible parameter for the screening of the cord blood unit (CBU) and the minimum cutoff of five NRBC/100 leukocytes can be a strategy in conjunction with the TNC to identify better units for cord blood bank sustainability.

Influence of hemolysis on nucleated red blood cells count.

Hemolysis is a common pre-analytical factor that can influence test results. Here, we explored the influence of hemolysis on nucleated red blood cells (NRBCs) count and tried to illustrate the mechanisms underlying this interference. From July 2019 to June 2021, 20 preanalytical hemolytic peripheral blood (PB) samples from inpatient at Tianjin Huanhu Hospital were evaluated using Sysmex XE-5000 automated hematology analyzer. When NRBC enumeration was positive and a flag was triggered, a 200-cell differential count was performed by experienced technologists on microscopic review. When the manual count was inconsistent with automated enumeration, samples will be re-collected. Plasma exchange test was performed to verify the influence factors of hemolyzed samples and the mechanical hemolysis experiment mimicking hemolysis that might occur during blood collection was performed to illustrate the underlying mechanisms. Hemolysis led to false-positive NRBC count and the value of NRBC was positively correlated with the degree of hemolysis. Hemolysis specimen shared a common scatter diagram: a "beard" on WBC/ basophil (BASO) channel and a "blue scatter line" on immature myeloid information (IMI) channel. Lipid droplets were found above the hemolysis specimen after centrifugation. Plasma exchange experiment confirmed that these lipid droplets interfered with NRBCs count. Mechanical hemolysis experiment implied further that broken red blood cells (RBCs) released lipid droplets causing the false-positive NRBCs count. In the present study, we firstly found that hemolysis could lead to false-positive NRBCs enumeration, which was associated with lipid droplets released from broken RBCs during hemolysis.

Nucleated red blood cells as a potential biomarker for predicting spontaneous intestinal perforation in premature infants

Background: The specific etiology of spontaneous intestinal perforation (SIP) and necrotizing enterocolitis (NEC) remains elusive. Both can present acutely without antecedent signs and can be difficult to differentiate. Neonatal nucleated red blood cell (NRBC) counts are part of the routine admission and serial CBCs drawn on premature neonates. Elevated NRBC counts could represent relative intrauterine hypoxemia, a possible risk factor for the development of SIP or NEC. Methods: We compared premature neonates with SIP to premature neonates with NEC and controls, matched gestational age (GA) and birth weight (BW). Kruskal-Wallis, Chi-Square, or Fisher's exact tests and univariate and multivariate nominal logistic regression models were used to estimate the association of baseline NRBC. Median times to SIP and NEC were calculated using the Kaplan-Meier method. All analyses were performed with SAS 9.4. Results: Male sex (72.5%), lower GA [Median (Q1, Q3): 25.1 (23.8, 28)], and lower BW [Median (Q1, Q3): 690 g (585, 1072)] had the highest incidence of SIP compared to NEC or controls. Increased baseline NRBC was associated with lower odds of developing NEC compared to controls [Median (Q1, Q3): 9 (5, 29) vs 19 (10, 51); OR (CI) 0.70 (0.55, 0.89), p-value = 0.0033]. Increased baseline NRBC was associated with higher odds of developing SIP compared to NEC [Median (Q1, Q3): 9 (5, 29) vs 19 (10, 51); OR (CI) 1.61 (1.18, 2.20) p-value = 0.0027]. There were no significant differences between intrauterine growth restriction (IUGR), maternal hypertension, chorioamnionitis, multiple births, or depressed APGAR scores in all three groups. NRBC for SIP neonates remained significantly higher at the day of life (DOL) 1-3 compared to other groups [Median (Q1, Q3): 23 (6, 93), p-value = 0.0020]. The percentage of patients with NRBC &gt;4, remained elevated for patients with SIP as late as week three (p = 0.0023). Conclusion: ELBW, male sex, and elevated baseline NRBC were significantly associated with the risk of developing SIP compared to NEC or controls. NRBC remained significantly elevated on DOL 1-3, compared to NEC or controls. Between the groups, there were no significant differences in perinatal stressors.

The Optimal Time of Peripheral Blood Hematopoietic Stem Cell Collection for Autologous Transplantation May Be Predicted By Immature Granulocytes and Nucleated Red Blood Cells

The time of initiating stem cell collection is very important for successful transplantation and is determined according to peripheral blood CD34+ cell count analysed by flow cytometry. In this study, we aimed to find the role of new complete blood count parameters to determine the optimal time of stem cell collection. Eighty-six patients who underwent stem cell mobilization were included in the study. Peripheral blood CD34+ cells and complete blood count were analysed on the same day. Patients with peripheral blood CD 34+ cell counts ≤ 20/μL and &amp;gt; 20/μL were determined as Group 1 and Group 2, respectively. The difference of CBC parameters between 2 groups and the relationship of these parameters with peripheral blood CD34+ cell counts were evaluated. It was found that immature granulocytes ratio, nucleated red blood cells ratio and count were positively correlated with peripheral blood CD34+ cell count and these parameters were significantly different between two groups (P&amp;lt; 0.001, P= 0.011 and P= 0.012, respectively). The new index formulated by using immature granulocytes and nucleated red blood cells ratios was significantly different between two groups (P&amp;lt;0.001), and had the highest diagnostic accuracy to determine the time of stem cell collection (AUC= 0.766). Immature granulocytes and nucleated red blood cells can be used to determine the time of peripheral blood stem cell collection faster and cheaper compared to flow cytometry.