Quantification of fetomaternal hemorrhage: a comparative study of the manual and automated microscopic Kleihauer-Betke tests and flow cytometry in clinical samples

Pelikan, Denise M.; Scherjon, Sicco A.; Mesker, Wilma E.; Groot-Swings, Godelieve M. de; Brouwer-Mandema, Geeske G.; Tanke, Hans J.; Kanhai, Humphrey H.H.

doi:10.1016/j.ajog.2004.01.007

Cited by 41 publications

(32 citation statements)

References 18 publications

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“…1.8 ml) FMH due to background fluorescence as previously reported [12] . In a recent study, FC was unable to distinguish the fluorescence signal of fetal blood cells from the background fluorescence of HbF contained in the adult cells [14] . Thus, the previous comparative studies of KBT and FC for FMH quantification have reported both over-and underestimation by KBT.…”

Section: Discussionmentioning

confidence: 99%

Assessment of Fetomaternal Hemorrhage by Flow Cytometry and Kleihauer-Betke Test in Rh-Negative Pregnancies

Savithrisowmya¹,

Singh

Kriplani³

et al. 2007

Gynecol Obstet Invest

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Objectives: To assess the efficacy of flow cytometry (FC) in the detection and quantification of fetomaternal hemorrhage (FMH) in comparison to the Kleihauer-Betke test (KBT). Methods: 25 unsensitized Rh-negative mothers who had delivered Rh-positive infants were included. Presence of FMH was determined by KBT and FC using FITC-labeled BRAD-3 antibodies. Results: FMH was detected in 19 (76%) patients by FC and 23 (92%) patients by KBT prior to delivery, and in 21 (84%) patients by FC and 23 (92%) patients by KBT after delivery. The mean volume of FMH in the post-delivery samples by KBT and FC were 0.34 ± 0.26 ml (range 0.05–1.2 ml) and 0.37 ± 0.57 ml (range 0.02–2.6 ml) respectively. The volume of post-delivery FMH estimated by KBT and FC correlated well (r = 0.75; ICC α = 0.73). A higher agreement between KBT and FC was seen in the 0.1–0.5 ml range (ĸ = 0.65; p < 0.01). Conclusions: Both manual KBT and FC using FITC-BRAD-3 antibodies show good sensitivity in detecting and quantifying fetal red cells. There is a good correlation between the methods in the 0.1- to 0.5-ml range of FMH.

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Section: Discussionmentioning

confidence: 99%

Assessment of Fetomaternal Hemorrhage by Flow Cytometry and Kleihauer-Betke Test in Rh-Negative Pregnancies

Savithrisowmya¹,

Singh

Kriplani³

et al. 2007

Gynecol Obstet Invest

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“…FCI was used to measure the rate of erythrocytes with a high expression of fetal hemoglobin (6)(7)(8)(9). For this assay, erythrocytes have to be isolated and made permeable.…”

Section: Quantification Of Fetal Erythrocytes By Fcimentioning

confidence: 99%

Significance of the volume of fetomaternal hemorrhage after performing prenatal invasive tests

Subirá

Uriel

Ma³

et al. 2010

Cytometry

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Background: Fetal erythrocytes cross the placenta during gestation, but invasive prenatal procedures might develop into fetomaternal hemorrhage (FMH). We examine whether flow cytometry immunophenotyping might be useful for measuring the volume of FMH after such procedures.Methods: Fetal erythrocytes (%) were determined in 153 pregnant women after amniocentesis (129) and chorionic villous sampling (24) using a monoclonal antibody against fetal hemoglobin. Fetal erythrocytes were identified for their high expression of fetal hemoglobin (HbF 11 ). Blood samples from two control groups, 53 healthy males and 21 pregnant women not submitted to invasive tests, were used to establish normal values of circulating HbF 11 erythrocytes in adults. Results: The highest percentage of HbF 11 erythrocytes in the control groups was 0.015%. The rate of HbF 11 erythrocytes in samples after invasive tests ranged between <0.01% and 0.15%. Seventy-three women (47%) had 0.015% HbF 11 erythrocytes, and this rate was higher in 80. Nine women presented >1 ml of FMH (volume of packed cells corresponding to 0.054-0.15% HbF 11 erythrocytes), but only two had sonographic evidence of bleeding.Conclusions: Most women in our series had a very low volume of FMH after the invasive tests. Acute bleeding should be thoroughly investigated in women with either more than 1 ml of packed cells or more than 0.05% of HbF 11 erythrocytes. Intermediate values between >0.015% and <0.05%, should be carefully considered depending on the week of gestation. Data obtained before 15 weeks might reflect previous cell trafficking between fetus and mother instead of acute hemorrhage. V C 2010 International Clinical Cytometry Society

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“…The Kleihauer-Betke test can be performed manually with stained blood smears or using a flow cytometry-based method and the technical merit and validation of the used method may be of influence on test performance. 28 Furthermore, fetal blood loss into the maternal intraperitoneal cavity, hence not detectable by the Kleihauer-Betke test, might well lead to RhD immunisation.…”

Section: Discussionmentioning

confidence: 99%

Risk Factors for RhD Immunisation Despite Antenatal and Postnatal Anti-D Prophylaxis

Koelewijn

Haas

Vrijkotte

et al. 2010

Obstetrical &Amp; Gynecological Survey

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Objective To identify risk factors for Rhesus D (RhD) immunisation in pregnancy, despite adequate antenatal and postnatal anti-D prophylaxis in the previous pregnancy. To generate evidence for improved primary prevention by extra administration of anti-D Ig in the presence of a risk factor.Design Case-control study.Setting Nation-wide evaluation of the Dutch antenatal anti-D-prophylaxis programme.Population Cases: 42 RhD-immunised parae-1, recognised by first-trimester routine red cell antibody screening in their current pregnancy, who received antenatal and postnatal anti-D Ig prophylaxis (gifts of 1000 iu) in their first pregnancy. Controls: 339 parae-1 without red cell antibodies.Methods Data were collected via obstetric care workers and/or personal interviews with women.Main outcome measure Significant risk factors for RhD immunisation in multivariate analysis.Results Independent risk factors were non-spontaneous delivery (assisted vaginal delivery or caesarean section) (OR 2.23; 95% CI:1.04-4.74), postmaturity ( ‡42 weeks of completed gestation: OR 3.07; 95% CI:1.02-9.02), pregnancy-related red blood cell transfusion (OR 3.51; 95% CI:0.97-12.7 and age (OR 0.89/year; 95% CI:0.80-0.98). In 43% of cases, none of the categorical risk factors was present.Conclusions In at least half of the failures of anti-D Ig prophylaxis, a condition related to increased fetomaternal haemorrhage (FMH) and/or insufficient anti-D Ig levels was observed. Hence, RhD immunisation may be further reduced by strict compliance to guidelines concerning determination of FMH and accordingly adjusted anti-D Ig prophylaxis, or by routine administration of extra anti-D Ig after a non-spontaneous delivery and/or a complicated or prolonged third stage of labour.

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Quantification of fetomaternal hemorrhage: a comparative study of the manual and automated microscopic Kleihauer-Betke tests and flow cytometry in clinical samples

Cited by 41 publications

References 18 publications

Assessment of Fetomaternal Hemorrhage by Flow Cytometry and Kleihauer-Betke Test in Rh-Negative Pregnancies

Assessment of Fetomaternal Hemorrhage by Flow Cytometry and Kleihauer-Betke Test in Rh-Negative Pregnancies

Significance of the volume of fetomaternal hemorrhage after performing prenatal invasive tests

Risk Factors for RhD Immunisation Despite Antenatal and Postnatal Anti-D Prophylaxis

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