Quantification of fetal nucleated cells in maternal blood of pregnant women with a male trisomy 21 fetus using molecular cytogenetic techniques

Krabchi, Kada; Gadji, Macoura; Samassékou, Oumar; Grégoire, Marie-Chantal; Forest, Jean‐Claude; Drouin, Régen

doi:10.1002/pd.1325

Cited by 25 publications

(33 citation statements)

References 27 publications

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“…This number is increased by 3 to 5 times in cases of Down's syndrome [4, 5]. Similar increases were also detected in different types of aneuploidies [4, 5]. According to our study, the number of detected fetal cells seemed to be underestimated by an average of 16% due to the occurrence of false-negative cells.…”

Section: Discussionsupporting

confidence: 83%

“…Using this methodology, we previously located a median of 2 to 6 fetal cells per milliliter of maternal blood in the second trimester of normal pregnancy between 18 and 22 weeks [9]. This number is increased by 3 to 5 times in cases of Down's syndrome [4, 5]. Similar increases were also detected in different types of aneuploidies [4, 5].…”

Section: Discussionmentioning

confidence: 99%

“…Low-frequency predictions were given by these studies that recognized fetal cells only by the presence of a Y chromosome signal in male pregnancy [4, 5, 9, 16, 17]. Although these studies yielded important information concerning the number of circulating fetal cells in maternal blood, the results are possibly skewed by the detection efficiency of these rare events by cumbersome and time-consuming manual scanning.…”

Section: Introductionmentioning

confidence: 99%

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Efficiency of Manual Scanning in Recovering Rare Cellular Events Identified by FluorescenceIn SituHybridization: Simulation of the Detection of Fetal Cells in Maternal Blood

Emad

Ayub²,

Samassékou³

et al. 2012

Journal of Biomedicine and Biotechnology

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Fluorescence in situ hybridization (FISH) and manual scanning is a widely used strategy for retrieving rare cellular events such as fetal cells in maternal blood. In order to determine the efficiency of these techniques in detection of rare cells, slides of XX cells with predefined numbers (1–10) of XY cells were prepared. Following FISH hybridization, the slides were scanned blindly for the presence of XY cells by different observers. The average detection efficiency was 84% (125/148). Evaluation of probe hybridization in the missed events showed that 9% (2/23) were not hybridized, 17% (4/23) were poorly hybridized, while the hybridization was adequate for the remaining 74% (17/23). In conclusion, manual scanning is a relatively efficient method to recover rare cellular events, but about 16% of the events are missed; therefore, the number of fetal cells per unit volume of maternal blood has probably been underestimated when using manual scanning.

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Section: Discussionsupporting

confidence: 83%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Efficiency of Manual Scanning in Recovering Rare Cellular Events Identified by FluorescenceIn SituHybridization: Simulation of the Detection of Fetal Cells in Maternal Blood

Emad

Ayub²,

Samassékou³

et al. 2012

Journal of Biomedicine and Biotechnology

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“…Of the abnormal cases, fetuses with trisomy 21 showed the highest mean number of NRBCs. This finding is consistent with previous studies (Bianchi et al ., 1997; Krabchi et al ., 2006; Mavrou et al ., 2007) that suggest more NRBCs are detected in maternal blood when the fetus is aneuploid, particularly trisomy 21. The maternal blood of pregnancies with trisomy 18 fetuses also had a higher number of NRBC/mL compared to trisomy 13 and triploid fetuses.…”

Section: Resultsmentioning

confidence: 99%

A microfluidics approach for the isolation of nucleated red blood cells (NRBCs) from the peripheral blood of pregnant women

et al. 2008

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Objective Nucleated red blood cells (NRBCs) have been identified in maternal circulation and potentially provide a resource for the monitoring and diagnosis of maternal, fetal, and neonatal health and disease. Past strategies used to isolate and enrich for NRBCs are limited to complex approaches that result in low recovery and less than optimal cell purity. Here we report the development of a high-throughput and highly efficient microfluidic device for isolating rare NRBCs from maternal blood. Material and Methods NRBCs were isolated from the peripheral blood of 58 pregnant women using a microfluidic process that consists of a microfluidic chip for size-based cell separation and a magnetic device for hemoglobin-based cell isolation. Results The microfluidic–magnetic combination removes nontarget red blood cells and white blood cells at a very high efficiency (∼99.99%). The device successfully identified NRBCs from the peripheral blood of 58/58 pretermination samples with a mean of 37.44 NRBC/mL (range 0.37–274.36 NRBC/mL). These results were compared with those from previous studies. Conclusion The microfluidic device results in an approximate 10- to 20-fold enrichment of NRBCs over methods described previously. The reliability of isolation and the purity of the NRBC product have the potential to enable the subsequent application of molecular diagnostic assays.

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“…One alternative way for obtaining information on the gestating fetus involves recovery of fetal material from maternal blood (6). Currently, two independent approaches have followed this direction, one aimed at the identification and analysis of fetal DNA (7)(8)(9)(10)(11)(12)(13); the other aimed at the identification of circulating fetal cells, including trophoblasts (14)(15)(16), nucleated fetal red blood cells (fNRBC) (17)(18)(19)(20), and nucleated white blood cells (21). Among these cell types, the most promising appears to be the identification and isolation of fNRBC and trophoblasts.…”

Section: Introductionmentioning

confidence: 99%

New trends in non-invasive prenatal diagnosis: Applications of dielectrophoresis-based Lab-on-a-chip platforms to the identification and manipulation of rare cells (Review)

Borgatti

Bianchi²,

Mancini³

et al. 2008

Int J Mol Med

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Abstract. The isolation of rare cells, such as fetal nucleated red blood cells and trophoblasts, from maternal blood for noninvasive prenatal diagnosis is a new field of research exhibiting several difficulties since this strategy requires unresolved basic technological protocols for a successful outcome. However, several achievements in the field of Laboratory-on-a-chip (Labon-a-chip) technology have provided clear advancements in projects aimed at the isolation of rare cells from biological fluids. Among the most interesting approaches are those based on dielectrophoresis (DEP). DEP-based Lab-on-a-chip platforms have been demonstrated to be suitable for several applications in biotechnology and biomedicine. DEP-based arrays are able to manipulate single cells, which can be identified and moved throughout the DEP chip to recovery places. DEP buffers are compatible with molecular interactions between monoclonal antibodies and target cells, allowing integration of these devices with magnetic cell sorting (MACS). DEP treatment does not alter the viability of manipulated cells.

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Quantification of fetal nucleated cells in maternal blood of pregnant women with a male trisomy 21 fetus using molecular cytogenetic techniques

Cited by 25 publications

References 27 publications

Efficiency of Manual Scanning in Recovering Rare Cellular Events Identified by FluorescenceIn SituHybridization: Simulation of the Detection of Fetal Cells in Maternal Blood

Efficiency of Manual Scanning in Recovering Rare Cellular Events Identified by FluorescenceIn SituHybridization: Simulation of the Detection of Fetal Cells in Maternal Blood

A microfluidics approach for the isolation of nucleated red blood cells (NRBCs) from the peripheral blood of pregnant women

New trends in non-invasive prenatal diagnosis: Applications of dielectrophoresis-based Lab-on-a-chip platforms to the identification and manipulation of rare cells (Review)

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