Feasibility of manual white blood cell counts as a predictor of neonatal sepsis in a low-resource setting
Christian N GoldingFrederik Schaltz-BuchholzerLilica SancaClara Clipet-JensenChristine Stabell BennNicholas AuKate ChipperfieldTobias R. KollmannNelly Amenyogbe
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Abstract Background Manual white blood cell (WBC) differential counts as a predictor for neonatal sepsis development in a low-resource setting have not been thoroughly evaluated. We hypothesized that manual differentiation (specifically immature:total [I:T] neutrophil ratios) would be feasible and useful as an adjunct to predict early-onset neonatal sepsis (EONS). Secondarily, we hypothesized that vaccination with bacillus Calmette-Guérin (BCG) and oral polio vaccine (OPV) could alter WBC differential counts and thus might reduce its predictive performance. Methods We performed a prospective cohort study within a randomized trial, randomizing healthy, high-risk newborns admitted to the nursery at the national hospital in Guinea-Bissau 1:1 to BCG+OPV at admission or at discharge (usual practice). Thin capillary blood films were prepared at 2 d of age in a subset of 268 neonates. WBC counts were assessed by microscopy and neonates were followed up for sepsis development within 2 weeks. Results Ninety-eight percent (264/268) of smears provided interpretable reads. Of the 264 children, 136 had been randomized to receive BCG+OPV prior to sampling; the remaining 128 were vaccinated at discharge. The I:T ratio (average 0.017) was lower among children who did not develop clinical sepsis but did not predict sepsis (p=0.70). Only three children had an I:T ratio >0.2 (associated with a higher probability of clinical sepsis in previous studies) but did not develop sepsis. Immunization did not alter WBC composition. Conclusions Manual WBC differentials are feasible in low-resource settings. WBC differentials are not affected by standard newborn immunization. However, the I:T ratio had no value in predicting subsequent development of sepsis.Keywords:
Neonatal Sepsis
White blood cell
Abstract Background: Neonatal sepsis remains an important cause of morbidity and mortality. The ability to quickly and accurately diagnose neonatal sepsis based on clinical assessments and laboratory blood tests remains difficult, where haemoculture is the gold standard for detecting bacterial sepsis in blood culture. It is also very difficult to study because neonatal samples are lacking. Methods : Forty-eight newborns suspected of sepsis admitted to the Neonatology Department of the Mother-Child Specialized Hospital of Tlemcen. From each newborn, a minimum of 1-2 ml of blood was drawn by standard sterile procedures for blood culture. The miRNA-23b level in haemoculture was evaluated by RT-qPCR. Results : miR-23b levels increased in premature and full-term newborns in early onset sepsis ( p < 0.001 and p < 0.005 respectively), but lowered in late onset sepsis in full-term neonates ( p < 0.05) compared to the respective negative controls. miR-23b levels also increased in late sepsis in the negative versus early sepsis negative controls ( p < 0.05). miR-23b levels significantly lowered in the newborns who died from both sepsis types ( p < 0.0001 and p < 0.05 respectively). In early sepsis, miR-23b and death strongly and negatively correlated (correlation coefficient = -0.96, p = 0.0019). In late sepsis, miRNA-23b and number of survivors (correlation coefficient = 0.70, p = 0.506) positively correlated. Conclusions : Lowering miR-23b levels is an important factor that favours sepsis development, which would confirm their vital protective role, and strongly suggest that they act as a good marker in molecular diagnosis and patient monitoring.
Neonatal Sepsis
Neonatology
Blood Culture
Positive correlation
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Background: Neonatal sepsis accounts for early-half of all neonatal death in India. Although the gold standard for the diagnosis of neonatal sepsis is by blood culture, there is a need for a test that is cheap, accurate, easily performed with quick availability of reports so that prompt and appropriate treatment is ensured. In this study we try to analyse the diagnostic value of I/T2ratio in comparison to I/T ratio and absolute neutrophil count (ANC) independently to correctly predict neonatal sepsis. Subjects and Methods: A time bound, prospective comparative study was done on neonates with suspected neonatal sepsis for a 2 year period. Neonates were then analysed in three groups depending on their blood culture results as no sepsis, probable sepsis and proven sepsis. I/T2was calculated for all the babies and an optimum cut off was found for the same. The diagnostic accuracy of I/T2 ratio with septic screen positivity and blood cultures were assessed. Results: 245 neonates included in the study were divided into three groups depending on whether they were blood culture positive sepsis, probable or clinical sepsis. The means of septic screen parameters including I/T2 were highly significant when compared between the three groups (p value <0.001). ROC curve created showed the AUC for I/T, I/T2 and ANC was 0.822, 0.809, 0.610 (p value <0.001) and a higher specificity. Conclusion: I/T2 shows promise as a parameter to screen for sepsis in neonates. Calculating the I/T2 ratio in conjunction with established septic screen parameters could enhance prediction of sepsis in newborns.
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Neonatal Sepsis
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Neonatal sepsis is the most common cause of mortality in newborns. Currently antibiotics and supportive care are the mainstay of treatment. Blood culture is considered as the gold standard for confirmation of diagnosis of neonatal sepsis. Here we have tried to develop a neonatal rat model of sepsis in order to better understand its progression. Lipopolysaccharide (LPS) is one of the common agents used to induce sepsis in rats. Here we found that LPS was ineffective in inducing sepsis in neonatal rats. We found that induction of live dose of Escherichia coli, one of the most common causes of neonatal sepsis was more effective than LPS injection. The rats were continuously monitored for the visual indications of sepsis development. Body weight, body temperature and the activity of rats were monitored continuously. Blood culture was done to check for the confirmation of diagnosis of sepsis. Further biochemical tests such as citrate, urease, indole and kliger-ion tests were done to confirm for E coli in the colonies of blood culture. The minimum effective dose of E coli needed to induce sepsis in neonatal rats was found to be 5*106 CFU of E coli.
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Neonatal sepsis can be classified into two subtypes depending upon whether the onset of symptoms is before 72 hours of life (early-onset neonatal sepsis-EONS) or later (late-onset neonatal sepsis-LONS). These definitions have contributed greatly to diagnosis and treatment by identifying which microorganisms are likely to be responsible for sepsis during these periods and the expected outcomes of infection. This paper focuses on the tools that microbiologist can offer to diagnose and eventually prevent neonatal sepsis. Here, we discuss the advantages and limitation of the blood culture, the actual gold standard for sepsis diagnosis. In addition, we examine the utility of molecular techniques in the diagnosis and management of neonatal sepsis.
Neonatal Sepsis
Gold standard (test)
Blood Culture
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Early diagnosis of neonatal sepsis continues to pose a problem to the clinician caring for newborns. Blood culture has been considered the gold standard for confirmation of diagnosis. An array of sepsis screen tests is now available for early diagnosis of neonatal sepsis. To study the role of sepsis screen tests in early diagnosis of neonatal sepsis. Blood cultures and sepsis screen tests were carried out on 200 neonates. The identification of the causative organism was carried out by standard identification tests. Sepsis screen was carried out with C‑reactive Protein, I/T ratio, Total leucocyte count, Absolute neutrophils count, Out of 200 neonates studied with clinical features of sepsis, 128 (64%) were blood culture positive. Among Sepsis screen tests, CRP was the most sensitive test (89.84%) with a specificity of 76.39%. I/T ratio, TLC and ANC showed sensitivity 76.56%, 44.53%, 32.81% and specificity 59.72%.76.39%, 81.94% respectively. Sepsis screen tests may help and can be used as an adjunct in the diagnosis. They are also useful in early detection of neonatal sepsis and can be used as prognostic indicators; thus helping to start early treatment with appropriate antibiotics.
Neonatal Sepsis
Blood Culture
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Abstract Background: Neonatal sepsis remains an important cause of morbidity and mortality. The ability to quickly and accurately diagnose neonatal sepsis based on clinical assessments and laboratory blood tests remains difficult, where haemoculture is the gold standard for detecting bacterial sepsis in blood culture. It is also very difficult to study because neonatal samples are lacking. Methods : Forty-eight newborns suspected of sepsis admitted to the Neonatology Department of the Mother-Child Specialized Hospital of Tlemcen. From each newborn, a minimum of 1-2 ml of blood was drawn by standard sterile procedures for blood culture. The miRNA-23b level in haemoculture was evaluated by RT-qPCR. Results : miR-23b levels increased in premature and full-term newborns in early onset sepsis ( p < 0.001 and p < 0.005 respectively), but lowered in late onset sepsis in full-term neonates ( p < 0.05) compared to the respective negative controls. miR-23b levels also increased in late sepsis in the negative versus early sepsis negative controls ( p < 0.05). miR-23b levels significantly lowered in the newborns who died from both sepsis types ( p < 0.0001 and p < 0.05 respectively). In early sepsis, miR-23b and death strongly and negatively correlated (correlation coefficient = -0.96, p = 0.0019). In late sepsis, miRNA-23b and number of survivors (correlation coefficient = 0.70, p = 0.506) positively correlated. Conclusions : Lowering miR-23b levels is an important factor that favours sepsis development, which would confirm their vital protective role, and strongly suggest that they act as a good marker in molecular diagnosis and patient monitoring.
Neonatal Sepsis
Blood Culture
Neonatology
Positive correlation
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Neonatal sepsis is a serious condition. Recent clinical studies have indicated that microRNAs (miRNAs) are key players in the pathogenesis of sepsis, which could be used as biomarkers for this condition.A total of 90 neonates with sepsis and 90 healthy neonates were enrolled in this study. qRT-PCR was performed to measure the expression levels of serum miR-34a-5p and miR-199a-3p.miR-34a-5p and miR-199a-3p serum levels were significantly reduced in neonates with sepsis compared with those in healthy neonates (P = 0.006 and P = 0.001, respectively). Significant correlations of miR-34a-5p and miR-199a-3p with each of TLC, RDW, RBS, and C-reactive protein (CRP) as well as SNAPII were observed, indicating their associations with the severity of neonatal sepsis.miR-34a-5p and miR-199a-3p may be useful as novel biomarkers in neonatal sepsis and may provide a new direction for its treatment.
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Background: Neonatal sepsis, a clinical syndrome of bacteremia with systemic signs and symptoms of infection in the first 4 weeks of life is a major cause of morbidity and mortality in newborn inborn. Early diagnosis is critical, as sepsis can progress more rapidly in neonates than in adults. An attempt was made to establish correlation between early neonatal sepsis screening & blood culture in neonates presenting with features of sepsis. The aim of this study is to assess the usefulness of sepsis screen in early diagnosis of neonatal septicemia.Materials and Methods: The study was done in Kist medical college and hospital, Nepal from October 2015 to October 2016. Statistical correlation between early indicators of sepsis screen & blood culture (considered as gold standard) was established in clinically suspicious cases of neonatal sepsis. Results: Out of 150 cases studied, 72 were culture positive. CRP (77.8%) and immature: total neutrophils ratio (73%) showed highest sensitivity. CRP (66.7%), I/T ratio (61.5%) and micro ESR (60.2%) showed highest specificity. Positive predictive value was highest for CRP (68.2%) followed by I/T ratio (63.8%) and corrected total leukocyte count (56.2%)Conclusion: Serum CRP is the most sensitive marker of sepsis. Use of peripheral smear study and CRP can be implicated effectively as a sepsis screen for early diagnosis of neonatal sepsis. The combination of parameters yielded better results than single tests and proved to be an invaluable tool for early diagnosis of neonatal sepsis.
Neonatal Sepsis
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Bacteremia
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Background: Neonatal septicaemia is one of the commonest causes of neonatal mortality and morbidity. Accurate and timely diagnosis of neonatal sepsis remains a major challenge to the pediatricians and neonatologists. In the present study, correlation between sepsis screening and blood culture in neonate presenting with features of sepsis is done to accelerate the diagnostic process and blood culture (considered gold standard) was evaluated as marker for sepsis detection and its effectiveness was compared with other septic markers.Methods: In present study, we emphasize to study early indicators of sepsis screen and their statistical correlation with blood culture (considered as gold standard).Results: As any sepsis screen parameters showed little correlation with blood culture, yet on combination it was found that specificity and positive predictive accuracy increased while sensitivity decreased them individual tests. Also combination of tests yield better results than single tests.Conclusions: The combination of sepsis makers yielded diagnostic results than single tests and proved to be an invaluable aid for early diagnosis of neonatal sepsis.
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Background Neonatal sepsis is a complex syndrome which is characterized by systemic inflammatory response. The high mortality rate of neonatal sepsis is contributed to late diagnosis and improper management. Aim of the Work The aim of this research is to examine the diagnostic utility of miR-34a in newborns with sepsis (who were newly diagnosed according to the clinical sepsis score and the hematology score of sepsis) and compare them to neonates without sepsis by using real-time PCR. Patients and Methods This study was conducted on 70 neonates from NICU of Ain Shams University hospital and permitted by Ain Shams ethical committee. Neonates were classified into two groups: 35 neonate with sepsis and 35 neonate without sepsis. In consideration, these neonates’ ages were ≥ 35 weeks, had no Congenital anomalies, Chromosomal abnormalities and had not been given any kind of antibiotics before obtaining blood specimens from them. Moreover, the study didn’ t include neonates with an inborn error of metabolism or any surgical procedures. Neonates without sepsis and neonates with sepsis were matched in terms of gender and age. In our study, blood samples of 2 ml were withdrawn from patients (within 72 hours from suspecting the diagnosis of neonatal sepsis) and from the control at matched gestation and postnatal ages, then patients were followed up. Results the obtained results showed that, the expression level of miR-34a was reduced by 5.2-fold in neonates with sepsis, with a P value of 0.001. Additionally, a negative correlation was detected between miR-34a expression level and either the clinical sepsis score (p = 0.163) and the hematology score (p = 0.122), and also a significant correlation was detected between the clinical sepsis and hematology scores in neonates with sepsis (p = 0.047). The study observed that, the miR-34a could discriminate neonates with clinical sepsis score, the biomarker sensitivity and specificity were 80% and 70%, respectively. In addition, the calculated sensitivity and specificity of miR-34a to discriminate neonates with hematology score were 85% and 87%; respectively. The cut-off value was <0.04 . Conclusion Based on the obtained results, there is a significant difference between the serum expression level of miR-34a and either the clinical sepsis score and hematology score . the lower expression level of miR-34a was detected in neonates with CSS >10. Similarly, the lower expression level of miR-34a was detected in neonates with high hematology score >4.
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Hematology
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