Shelley M Lawrence1, Jeffrey Eckert2, Marjorie Makoni2, H Anne Pereira3. 1. Department of Pediatrics, Section of Neonatal-Perinatal Medicine, University of Oklahoma Health Sciences Center, College of Medicine, Oklahoma, USA slawrence@ucsd.edu. 2. Department of Pediatrics, Section of Neonatal-Perinatal Medicine, University of Oklahoma Health Sciences Center, College of Medicine, Oklahoma, USA. 3. Department of Pharmaceutical Sciences, University of Oklahoma Health Sciences Center, College of Pharmacy, Oklahoma, USA.
Abstract
BACKGROUND: Automated complete blood counts (CBCs) provide imprecise measures of neonatal neutrophil composition due to the presence of nucleated red cells, which masquerade as white cells. Manual differentials (MDs) must be performed but can be flawed due to technician inexperience or limited cell counts. Our study evaluated whether flow cytometry could more accurately determine neutrophil composition in term newborns compared with adult controls. METHODS: Neutrophils were obtained by venipuncture in adults (n=10) and via cord blood from neonates delivered vaginally (n=21) or by primary cesarean section (n=17). Samples were processed by both flow cytometry and CBC with MD and results compared. Flow cytometry findings were verified using gene expression analysis of granule proteins. RESULTS: Immature-to-total neutrophil ratios were used to signify neutrophil composition. Flow cytometry was superior to manual differentials due to its ability to identify the earliest neutrophil forms. Significant differences between newborns and adults were determined using flow cytometry (0.12 versus 0.05; p=0.01), but not MDs (p=0.18). Neonates had similar findings irrespective of labor and method used. Verification of neutrophil maturity was completed using gene expression analysis of granule proteins. CONCLUSIONS: These results raise concerns about the accuracy of using CBCs with MDs to determine neutrophil composition. Further investigation of flow cytometry in clinical practice is warranted.
BACKGROUND: Automated complete blood counts (CBCs) provide imprecise measures of neonatal neutrophil composition due to the presence of nucleated red cells, which masquerade as white cells. Manual differentials (MDs) must be performed but can be flawed due to technician inexperience or limited cell counts. Our study evaluated whether flow cytometry could more accurately determine neutrophil composition in term newborns compared with adult controls. METHODS: Neutrophils were obtained by venipuncture in adults (n=10) and via cord blood from neonates delivered vaginally (n=21) or by primary cesarean section (n=17). Samples were processed by both flow cytometry and CBC with MD and results compared. Flow cytometry findings were verified using gene expression analysis of granule proteins. RESULTS: Immature-to-total neutrophil ratios were used to signify neutrophil composition. Flow cytometry was superior to manual differentials due to its ability to identify the earliest neutrophil forms. Significant differences between newborns and adults were determined using flow cytometry (0.12 versus 0.05; p=0.01), but not MDs (p=0.18). Neonates had similar findings irrespective of labor and method used. Verification of neutrophil maturity was completed using gene expression analysis of granule proteins. CONCLUSIONS: These results raise concerns about the accuracy of using CBCs with MDs to determine neutrophil composition. Further investigation of flow cytometry in clinical practice is warranted.
Authors: B C MacQueen; R D Christensen; B A Yoder; E Henry; V L Baer; S T Bennett; H M Yaish Journal: J Perinatol Date: 2016-06-09 Impact factor: 2.521
Authors: Marjorie Makoni; Jeffrey Eckert; H Anne Pereira; Victor Nizet; Shelley M Lawrence Journal: Early Hum Dev Date: 2016-07-16 Impact factor: 2.079
Authors: Eduarda Cristina Martins; Lilian da Fe Silveira; Karin Viegas; Andrea Diez Beck; Geferson Fioravantti Júnior; Rafael Viegas Cremonese; Priscila Schmidt Lora Journal: Rev Bras Ter Intensiva Date: 2019-03-21