Validation of automatic scanning of microscope slides in recovering rare cellular events: application for detection of fetal cells in maternal blood

Emad, Ahmed; Bouchard, Éric F.; Lamoureux, Josée; Ouellet, Annie; Dutta, Aparajita; Klingbeil, Uli; Drouin, Régen

doi:10.1002/pd.4345

Cited by 25 publications

(28 citation statements)

References 40 publications

(93 reference statements)

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“…Attempts to recover male fnRBCs in blood samples obtained prior to CVS or pregnancy termination from women carrying male pregnancies failed in 60–70% of cases leaving some doubt as to whether this cell type is present in sufficient numbers for routine analysis during the first trimester. In contrast, two groups have demonstrated that there are one to six fetal cells per milliliter of mother's blood during the first trimester using very reliable methods for Y chromosome FISH, and showed that these cells are certainly or most likely trophoblasts. Based on these reports, we have focused exclusively on detecting trophoblasts.…”

Section: Introductionmentioning

confidence: 99%

Evidence for feasibility of fetal trophoblastic cell‐based noninvasive prenatal testing

et al. 2016

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ObjectiveThe goal was to develop methods for detection of chromosomal and subchromosomal abnormalities in fetal cells in the mother's circulation at 10–16 weeks' gestation using analysis by array comparative genomic hybridization (CGH) and/or next‐generation sequencing (NGS).MethodNucleated cells from 30 mL of blood collected at 10–16 weeks' gestation were separated from red cells by density fractionation and then immunostained to identify cytokeratin positive and CD45 negative trophoblasts. Individual cells were picked and subjected to whole genome amplification, genotyping, and analysis by array CGH and NGS.ResultsFetal cells were recovered from most samples as documented by Y chromosome PCR, short tandem repeat analysis, array CGH, and NGS including over 30 normal male cells, one 47,XXY cell from an affected fetus, one trisomy 18 cell from an affected fetus, nine cells from a trisomy 21 case, three normal cells and one trisomy 13 cell from a case with confined placental mosaicism, and two chromosome 15 deletion cells from a case known by CVS to have a 2.7 Mb de novo deletion.ConclusionWe believe that this is the first report of using array CGH and NGS whole genome sequencing to detect chromosomal abnormalities in fetal trophoblastic cells from maternal blood. © 2016 The Authors. Prenatal Diagnosis published by John Wiley & Sons, Ltd.

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Section: Introductionmentioning

confidence: 99%

Evidence for feasibility of fetal trophoblastic cell‐based noninvasive prenatal testing

et al. 2016

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“…A recent study published by Drouin et al looked at the validation of automatic scanning of microscopic slides to detect fetal cells in maternal blood [56]. The frequency of fetal cells per 1 ml of maternal blood identified by this approach ranged from 3 to 6 cells in normal pregnancies and 13 to 21 cells in Down syndrome pregnancies.…”

Section: Type and Number Of Fetal Cells In The Maternal Circulationmentioning

confidence: 98%

Cell-free DNA versus intact fetal cells for prenatal genetic diagnostics: what does the future hold?

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Feinberg

Wapner

et al. 2015

Expert Review of Molecular Diagnostics

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Prenatal screening and diagnosis is currently focused on the development of a noninvasive prenatal diagnostic test capable of detecting abnormalities similar to those attainable with an invasive test. One contender is cell-free fetal DNA circulating in maternal plasma and the other is intact fetal cells either from the maternal blood or the cervix. Once adequate fetal DNA is available, laboratory analytic techniques, such as sequencing and microarray, can be applied allowing detection of most cytogenetic and Mendelian fetal disorders. The question is: how close are we to achieving this feat, and what does the future hold?

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“…Our findings provide a proof for the feasibility of NIPD from very small numbers of fetal cells and suggest that detection, amplification and analysis of as few as 5 fetal cells that could be obtained from 2-3 ml of maternal peripheral blood [45] can sufficient enough to provide an accurate NIPD. The few fetal cells required together with the use of automatic scanning for the detection of fetal cells [73] and SCs-WGA will help to overcome the problem of rarity of these cells in the maternal blood and make their use in NIPD much more easily achievable. Although this method is not yet completely mature due to the absence of a perfect antigen that can recognize 100% of fetal cells, relentless efforts continue and should lead to the development of this antigen in the near future [74,75].…”

Section: Discussionmentioning

confidence: 99%

“…QF-PCR, which is less expensive and almost entirely automated, enables more women to undergo prenatal diagnosis without a significant increase in health expenditure [37,38,71,72]. The use of innovative technology like automatic cellular scanning of slides for the detection of fetal cells from maternal peripheral blood, which we have recently optimized and used for detection and quantification of fetal cells in the maternal blood in euploid and aneuploid pregnancies, will help to avoid the traditional cumbersome and time-consuming manual detection of rare fetal cells from maternal blood [73]. …”

Section: Discussionmentioning

confidence: 99%

Rapid Aneuploidy Detection of Chromosomes 13, 18, 21, X and Y Using Quantitative Fluorescent Polymerase Chain Reaction with Few Microdissected Fetal Cells

et al. 2015

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Objectives: Analysis of DNA from small numbers of cells, such as fetal cells in maternal blood, is a major limiting factor for their use in clinical applications. Traditional methods of single-cells whole genome amplification (SCs-WGA) and accurate analysis have been challenging to date. Our purpose was to assess the feasibility of using a few fetal cells to determine fetal sex and major chromosomal abnormalities by quantitative fluorescent polymerase chain reaction (QF-PCR). Methods: Cultured cells from 26 amniotic fluid samples were used for standard DNA extraction and recovery of 5 fetal cells by laser-capture microdissection. SCs-WGA was performed using the DNA from the microdissected cells. PCR amplification of short tandem repeats specific for chromosomes 13, 18, 21, X and Y was performed on extracted and amplified DNA. Allele dosage and sexing were quantitatively analyzed following separation by capillary electrophoresis. Results: Microsatellite QF-PCR analysis showed high concordance in chromosomal copy number between extracted and amplified DNA when 5 or more cells were used. Results were in concordance with that of conventional cytogenetic analysis. Conclusion: Satisfactory genomic coverage can be obtained from SCs-WGA. Clinically, SCs-WGA coupled with QF-PCR can provide a reliable, accurate, rapid and cost-effective method for detection of major fetal chromosome abnormalities.

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Validation of automatic scanning of microscope slides in recovering rare cellular events: application for detection of fetal cells in maternal blood

Cited by 25 publications

References 40 publications

Evidence for feasibility of fetal trophoblastic cell‐based noninvasive prenatal testing

Evidence for feasibility of fetal trophoblastic cell‐based noninvasive prenatal testing

Cell-free DNA versus intact fetal cells for prenatal genetic diagnostics: what does the future hold?

Rapid Aneuploidy Detection of Chromosomes 13, 18, 21, X and Y Using Quantitative Fluorescent Polymerase Chain Reaction with Few Microdissected Fetal Cells

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