Detection of fetal red cells in fetomaternal hemorrhage using a fetal hemoglobin monoclonal antibody by flow cytometry

Abstract

The anti-HbF flow cytometric method for detection of fetal cells offers a simple, reliable, and more precise alternative to the Kleihauer-Betke manual technique for the assessment of fetomaternal hemorrhage. The method has additional potential applications for the study of HbF levels or frequency of adult red cells with low levels of HbF (F cells) in individuals with hemoglobinopathies.

Evaluation of FMH QuikQuant for the detection and quantification of fetomaternal hemorrhage

“…However, this assay lacks specificity and exhibits a large inter-observer variation resulting in potential overtreatment risks related to plasma-derived products and unnecessary treatment cost (20)(21)(22). In accordance with previous studies, we reported an overestimation of FMH by the KBT (12,15,23). The KBT presents stained maternal F cells that may be counted as fetal RBCs, leading to these false positive results (20,23,24).…”

“…We found a coefficient of variation lower than 20% for values higher than 10 fetal RBCs per 10,000 maternal RBCs (0.1%) which is better than the KBT and in accordance with previous studies (4,13). Additionally, the kit design includes a positive marker of nucleated cells, thus facilitating exclusion of these interfering cells with autofluorescence levels similar in fluorescein isothiocyanate (FITC) fluorescence intensity as anti-HbF labeled fetal RBC (12,27). We adapted the gating strategy to enhance the fetal RBCs discrimination.…”

“…However, such strategy is limited to the determination of anti-RhD immunoglobulin dose, as it cannot distinguish fetal RBCs from adult cells in cases of RhD compatibility. In the last decade, FCM approaches using intracellular detection of HbF have been developed (4,(12)(13)(14)(15)(16)(17). Monoclonal anti-HbF antibody allows discrimination of three distinct populations: fetal RBCs, F cells, and adult RBCs (12,14) The quantification of F cells provided by FCM eliminates a major drawback of KBT.…”

“…In the last decade, FCM approaches using intracellular detection of HbF have been developed (4,(12)(13)(14)(15)(16)(17). Monoclonal anti-HbF antibody allows discrimination of three distinct populations: fetal RBCs, F cells, and adult RBCs (12,14) The quantification of F cells provided by FCM eliminates a major drawback of KBT. In this context, FMH QuikQuant is a new CE marked kit including a monoclonal anti-HbF antibody and propidium iodide (PI) as a specific marker of nucleated cells (18,19).…”

Detection of weak D and DVI red cells in D‐negative mixtures by flow cytometry: implications for feto‐maternal haemorrhage quantification and D typing policies for newborns

“…Recently, the use of anti-fetal haemoglobin antibodies for the detection of fetal cells by flow cytometry has been described (Davis et al, 1998 (Scott, 1996).…”