​Nucleated Red Blood Cell Count as Earliest Prognostic Marker for Adverse Neonatal Outcome in Neonatal Sepsis

Introduction: The most common cause of neonatal mortality is neonatal sepsis, the diagnosis of which depends on bloodculture, which has low sensitivity and takes time. We hypothesize that demonstration of elevated NRBC levels in neonatalsepsis might help in predicting an adverse neonatal outcome.Aim of the Study: To analyse elevated NRBC it can serve as a prognostic marker for neonatal sepsis and an increased riskfor adverse neonatal outcome.Materials and Methods: This is a hospital based prospective study done in neonates who are admitted in NICU of MaduraiMedical College with risk factors or clinical features of sepsis. After getting informed consent, the maternal details and examinationfindings were recorded and blood sample taken for sepsis screen, blood culture and peripheral smear for NRBC.Results: The sensitivity of NRBC in identifying sepsis was 81.5%, its specificity was 61.76%, positive predictive value was 70.4%and negative predictive value was 75%. In the neonates who expired, serial NRBC counts (mean – 22.4) were significantlyincreased from baseline value (mean 17.3).Conclusion: NRBC is significantly elevated in the neonatal sepsis and is a predictor of adverse neonatal outcome.

Nucleated RBCs in cord blood as a predictor of fetal asphyxia

Background: Perinatal asphyxia is a major cause of acute mortality and chronic neurologic disability amongst survivors, and is a complication that occurs between 2-10% of deliveries. No single parameter can define perinatal asphyxia, rather a combination of parameters like fetal distress, meconium-stained liquor, low Apgar score, umbilical cord blood pH and clinical features of hypoxicischemic encephalopathy (HIE) can predict it. Aim: To study the correlation between nucleated red blood cell count (NRBC), fetal acidosis and clinical markers of asphyxia. Materials and methods: 52 pregnant women with fetal distress as a study group and 51 pregnant women without distress as a control the ed group were selected and their cord blood was analyzed for pH and nucleated RBCs. The results were analyzed. Result: Higher NRBC count was detected in the study group with signs of asphyxia (Thick MSAF, Non-Reassuring FHR Pattern, Low APGAR score). Conclusion: NRBC count is a simple bedside test to diagnose fetal asphyxia. Since it is cost-effective and does not require any special expertise or any high-tech facilities, it may be a useful, reliable, inexpensive and easily available markan er to evaluate perinatal asphyxia, especially in a resourcepoor country like ours, where blood gas analysis facilities are not available in the majority of place.

Evaluation of the Nucleated Red Blood Cells Count of Sysmex XN-9000 Automatic Hematology Analyzer and the Validation of Manual Method / 现代检验医学杂志

Objective To evaluate the nucleated red blood cell (NRBC)count of Sysmex XN-9000 automatic hematology anal-ysis lines comparing with manual method,and verify the accuracy of the analyzer results.Methods 60 blood samples with more than 1% of NRBCs detected by XN-9000 were counted NRBCs by traditional manual microscopy in blood smears,and verified the analyzer results.Results According to the reliability analysis,the results of total 60 samples were all within the range of the reliability;correlation analysis showed that correlation coefficient (r)of group NRBC (%)1~10 and>10 were 0.972 1 and 0.996 2,respectively.There were no significant differences between them (P>0.05).Conclusion Compared with manual method,the results of NRBC count of XN-9000 were within the range of reliability,and showed good correla-tion.The analyzer test results of NRBC were accurate and reliable and could be applied to the detection of clinical samples.

Evaluation of the Measurement of Nucleated Red Blood Cells in Neonatal Blood with ADVIA 2120 / 임상검사와정도관리

BACKGROUND: Nucleated red blood cells (NRBCs) are immature erythrocytes. The current line of thought is that increased NRBC production in the immediate neonatal state primarily reflects hypoxic injury and a direct response to mediators of inflammation in newborns with early-onset neonatal sepsis. So, the accurate, reproducible, and timely reporting of NRBC is an important function of the clinical laboratory. The aim of this study is to evaluate the performances of the ADVIA 2120 Hematology System (Siemens, USA) in the NRBCs measurement and compare them with the conventional microscopic method. METHODS: The study was performed in Kangnam Sacred Heart Hospital from March to September, 2009. One hundred twenty neonatal samples were submitted to measure NRBC count. For comparison, 225 blood samples except neonates were also included. The specimens were analyzed for the NRBC counts both with ADVIA 2120 analyzer and by microscopic method. RESULTS: Correlations of NRBC counts form the ADVIA 2120 compared with microscopic results were excellent in neonatal and in all specimens, r=0.8630 and r=0.9012, respectively. Sensitivity and specificity for the presence of NRBC was 98.2% and 94.2%, respectively. Concordance rate was 95.5%. There were some discordant results in cases of chronic lymphocytic leukemia and iron deficient anemia. CONCLUSIONS: The ADVIA 2120 NRBC method correlated well with the manual microscopic method. The automatic correction of the WBC count and differential, the shorter turnaround time compared to microscopic counting are additional advantages of automatic method. The automatic NRBC counting is considered to be a useful method in neonatal hematology.

Prenatal genetic diagnosis from maternal blood: simultaneous immunophenotyping and FISH of fetal nucleated erythrocytes isolated by negative and positive magnetic activated cell sorting

Fetal nucleated red blood cells (nRBCs) are rare in maternal circulation, but their presence constitutes a potential source of non-invasive prenatal genetic diagnosis. This study was undertaken to establish a non-invasive prenatal genetic diagnosis method using isolated fetal nRBCs. A multi-step method including triple density gradient and magnetic activated cell sorting (MACS) using CD45 and CD71, cytospin centrifugation, K-B staining, and glycophorin A-immuno fluorescence in situ hybridization (GPA-immuno FISH) was performed. The study population included 65 patients from 8 to 41 weeks of gestation, and fetal nRBC was separated from all cases. The number of fetal nRBCs retrieved was 12.8 +/- 2.7 in 8 to 11 gestational weeks, 15.2 +/- 6.5 in 12 to 18 gestational weeks, 16.4 +/- 6.5 in 19 to 23 gestational weeks, 10.6 +/- 3.2 in 24 to 28 gestational weeks, and 5.5 +/- 1.9 in 35 to 41 gestational weeks: the mean number of nRBCs collected from 20 ml of maternal peripheral blood was 13.7 +/- 6.2. The highest value of yield was 45.6% from 12 to 18 weeks gestation. The fetal sex determination confirmed by amniocentesis or chorionic villus sampling showed 100% sensitivity and 91.7% specificity for males; 91.7% sensitivity and 100% specificity for females. We showed that fetal cells can be reliably enriched from maternal blood and that they can be used for detecting specific chromosomes by FISH with a specificity superior to current non-invasive methods.

Predicting Intraventricular Hemorrhage by the Nucleated Erythrocytes Counts in Preterm Neonates

PURPOSE: Prolonged fetal hypoxia stimulates erythropoiesis in fetal life and induces increased nucleated erythrocytes(NRBC) counts at the early newborn period. To evaluate the relationship between prolonged fetal hypoxia and neonatal intraventricular hemorrhage (IVH), and the prediction of neonatal IVH by neonatal NRBC. METHODS: We compared the daily courses of the absolute NRBC count in preterm new- boms at 34 weeks' gestation or earlier with(n=17) and without(n=20) IVH for 7 days of life. RESULTS: Absolute NRBC counts at birth were higher in neonates with IVH than in control neonates(2,499/mm3+/-3,748 and 412/mm3+/-272, respectively, P=0.0022). The cut-off value of 1,000/mm3 for absolute NRBC counts at birth showed the best parameter estimate of the predictive model for IVH at early newborn period with 100% of positive predictive value and 74.1% of negative predictive value. CONCLUSION: Prolonged fetal hypoxia inducing fetal erythropoiesis near labor is closely related to IVH at early newborn period. Thabsolute NRBC counts at birth is the very important predictable marker for the condition.

Analysis of Fetal Nucleated Erythrocytes in Maternal Blood by Fluorescence In Situ Hybridization(FISH) / 대한산부인과학회잡지

The isolation of fetal cells from maternal circulation has the potential to allow relativelyself prenatal diagosis for all pregnant women. The present technology, however, has notreached the accuracy required for clinical diagnosis because of maternal cell contaminationSo we published a new method for enrichment of nRBC in a fetal cell isolation(1996).In this study, attempted to FISH analysis of nRBC which was isolated by our ownmethods. We evaluated the efficiency of FISH.As the results, we have successfully used FISH on enriched nRBC.We were able to identified 2 abnormal fetus which were confirmed by conventionalcytogenentic study as Down syndrome(Fig.1) and Klinefeltre syndrome(Fig.2). And thesensitivity and specificity for FISH was 86%(49/57) and 92.3%(36/39), respectively.According to our results, fetal cell analysis by FISH can be reliable used for prenatalaneuploidy diagnosis. However, the problems of enrichment of the fetal cell and FISH probeor condition should be over come before analyze.