Detection of Fetal Trisomies 21 and 18 From Maternal Blood Using Triple Gradient and Magnetic Cell Sorting

Gänshirt‐Ahlert, Dorothee; Börjesson‐Stoll, Regina; Burschyk, Monika; Dohr, Angelika; Garritsen, Henk; Helmer, Elisabeth; Miny, Peter; Velasco, Marta; Walde, Corinna; Patterson, David; Teng, Nelson N.H.; Bhat, Neelima M.; Bieber, Marcia M.; Holzgreve, Wolfgang

doi:10.1111/j.1600-0897.1993.tb00620.x

Cited by 178 publications

(146 citation statements)

References 12 publications

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“…Our findings are at variance to those of Gänshirt-Ahlert et al 12,13 , who also used triple density gradient centrifugation and anti-CD71 magnetic cell sorting to enrich fetal cells in maternal blood and reported no significant difference in the number of erythroblasts between pre-CVS and post-CVS samples. In contrast, Jansen et al 14 , who isolated fetal cells by Ficoll single density centrifugation and maternal depletion with CD45 and CD14 and CD71 magnetic cell sorting enrichment and then applied FISH for the Y-chromosome, from blood samples obtained immediately before and 5 -20 minutes after CVS, found an increase in the number of fetal cells in 10 out of 19 male pregnancies examined.…”

Section: Discussioncontrasting

confidence: 56%

“…In contrast, Jansen et al 14 , who isolated fetal cells by Ficoll single density centrifugation and maternal depletion with CD45 and CD14 and CD71 magnetic cell sorting enrichment and then applied FISH for the Y-chromosome, from blood samples obtained immediately before and 5 -20 minutes after CVS, found an increase in the number of fetal cells in 10 out of 19 male pregnancies examined. Possible reasons for the difference in findings in the studies of Gänshirt-Ahlert et al 12,13 are that firstly, the pre-CVS and post-CVS blood samples were obtained from two different groups of women; secondly, the range of gestational age of the pregnancies examined was not indicated, particularly in relation to the similarity of gestation of the two groups, and previous studies have shown a biological variation in the number of fetal cells present in maternal blood at different gestations 17 -19 ; and thirdly, the distribution of normal and abnormal karyotype in the pregnancies examined before and after CVS is not given and it is known that in trisomic pregnancies the number of fetal cells in maternal blood is increased 6,7 .…”

Section: Discussionmentioning

confidence: 73%

“…The median gestation at CVS was 13 (range [11][12][13][14] weeks and at termination was 14 (range 12-15) weeks.…”

Section: Resultsmentioning

confidence: 99%

“…There is also some controversial evidence that invasive procedures may cause an increase in the number of fetal cells enriched from maternal blood. Gänshirt-Ahlert et al 12,13 used triple density gradient and MACS to determine the total number of enriched erythroblasts before and after invasive procedures and reported no significant differences. In contrast, Jansen et al 14 , studied 19 pregnant women carrying male fetuses at 11 -14 weeks of gestation, and reported that CVS was associated with an increase in the number of fetal cells, using Y-signals on fluorescence in situ hybridisation (FISH) in enriched erythroblasts from maternal blood, in 10 of the 19 male pregnancies.…”

Section: Introductionmentioning

confidence: 99%

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Distribution of fetal erythroblasts in maternal blood after chorionic villous sampling

Al-Mufti

2003

BJOG: An International Journal of Obstetrics and Gynaecology

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Section: Discussioncontrasting

confidence: 56%

Section: Discussionmentioning

confidence: 73%

“…The median gestation at CVS was 13 (range [11][12][13][14] weeks and at termination was 14 (range 12-15) weeks.…”

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Distribution of fetal erythroblasts in maternal blood after chorionic villous sampling

Al-Mufti

2003

BJOG: An International Journal of Obstetrics and Gynaecology

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“…Many methods like Fluorescence Activated Cell Sorting (FACS), Magnetic Activated Cell Sorting (MACS), charge flow separation, immunofluorescent beads, lectins, etc. have been exploited for the isolation of fetal nucleated red blood cells (NRBCs) from maternal peripheral blood (1)(2)(3)(4)(5). Of these, FACS has been the most widely used technique.…”

mentioning

confidence: 99%

A comparison of the choice of monoclonal antibodies for recovery of fetal cells from maternal blood using FACS for noninvasive prenatal diagnosis of hemoglobinopathies

D’Souza

Ghosh

Colah

2008

Cytometry Part B Clinical

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Background: Isolation of adequate numbers of fetal cells circulating in the maternal circulation is the major hurdle in developing noninvasive prenatal diagnostic procedures. We used flow cytometry and a combination of different monoclonal antibodies to compare the yield and purity of the fetal nucleated red blood cells at different periods of gestation.Methods: Using a Percoll discontinuous gradient, the fetal nucleated erythrocytes were enriched from 7 ml maternal blood. In 100 samples, the enriched cells were stained with CD45, anti-fetal hemoglobin, and glycophorin A antibodies and in 30 samples they were stained with CD45, anti-fetal hemoglobin, and CD71 and then sorted and used for fetal diagnosis of hemoglobinopathies.Results: Using the first set of antibodies, although we were able to obtain a higher percentage of fetal nucleated red cells (0.07% 6 0.2%) as compared to the second set which yielded comparatively smaller numbers (0.025% 6 0.03%), there was some compromise in purity.Conclusion: Using CD45, anti-fetal hemoglobin and CD71 would be preferred as minimizing maternal contamination is more important than yield for prenatal diagnosis. q

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Investigation of maternal blood enriched for fetal cells: Role in screening and diagnosis of fetal trisomies

et al. 1999

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Prenatal diagnosis of chromosomal abnormalities relies on assessment of risk followed by invasive testing in the group with highest risk. Assessment of risk by a combination of maternal age and fetal nuchal translucency and invasive testing in the 5% of the population with the highest risk would identify about 80% of trisomy 21 pregnancies. Preliminary reports suggest that chromosomal abnormalities can also be diagnosed by fluorescent in situ hybridization (FISH) in maternal blood enriched for fetal cells. This study examines the potential role of this method on the prenatal diagnosis of fetal trisomies. Maternal blood was obtained before invasive testing in 230 pregnancies at 10-14 weeks of gestation. After enrichment for fetal cells, by triple density centrifugation and anti-CD71 magnetic cell sorting, FISH was performed and the proportion of cells with positive signals in the chromosomally normal and abnormal groups was determined. Fetal karyotype was normal in 150 cases and abnormal in 80 cases, including 36 with trisomy 21. Using a 21 chromosome-specific probe, three-signal nuclei were present in at least 5% of the enriched cells from 61% of the trisomy 21 pregnancies and in none of the normal pregnancies. For a cut-off of 3% of three-signal nuclei the sensitivity for trisomy 21 was 97% for a false positive rate of 13%. Similar values were obtained in trisomies 18 and 13 using the appropriate chromosome-specific probe. Examination of fetal cells from maternal blood may provide a noninvasive prenatal diagnostic test for trisomy 21 with the potential of identifying about 60% of affected pregnancies. Alternatively, this technique can be combined with maternal age and fetal nuchal translucency as a method of selecting the high-risk group for invasive testing. Potentially, 80% of trisomy 21 pregnancies could be identified after invasive testing in less than 1% of the pregnant population.

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Detection of Fetal Trisomies 21 and 18 From Maternal Blood Using Triple Gradient and Magnetic Cell Sorting

Abstract: Our new approach for noninvasive prenatal diagnosis has proven to be reliable in this first series.

Cited by 178 publications

References 12 publications

Distribution of fetal erythroblasts in maternal blood after chorionic villous sampling

Distribution of fetal erythroblasts in maternal blood after chorionic villous sampling

A comparison of the choice of monoclonal antibodies for recovery of fetal cells from maternal blood using FACS for noninvasive prenatal diagnosis of hemoglobinopathies

Investigation of maternal blood enriched for fetal cells: Role in screening and diagnosis of fetal trisomies

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