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

2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
Ahmed Emad ◽  
Seemi Ayub ◽  
Oumar Samassékou ◽  
Marie-Chantal Grégoire ◽  
Macoura Gadji ◽  
...  
Keyword(s):  
Efficient Method ◽  
Unit Volume ◽  
Detection Efficiency ◽  
Maternal Blood ◽  
Fetal Cells ◽  
Cellular Events ◽  
Probe Hybridization ◽  
Rare Cells

Fluorescencein situhybridization (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.

scholarly journals FISH Analysis of All Fetal Nucleated Cells in Maternal Whole Blood: Improved Specificity by the Use of Two Y-chromosome Probes

2005 ◽  
Vol 53 (3) ◽  
pp. 319-322 ◽  
Author(s):  
Susanne Mergenthaler ◽  
Tatiana Babochkina ◽  
Vivian Kiefer ◽  
Olaf Lapaire ◽  
Wolfgang Holzgreve ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Y Chromosome ◽  
Blood Cells ◽  
Maternal Blood ◽  
Fish Analysis ◽  
Blood Samples ◽  
Fetal Cells ◽  
Nucleated Red Blood Cells ◽  
Noninvasive Prenatal Diagnosis

Current cytogenetic approaches in noninvasive prenatal diagnosis focus on fetal nucleated red blood cells in maternal blood. This practice may be too restrictive because a vast proportion of other fetal cells is ignored. Recent studies have indicated that fetal cells can be directly detected, without prior enrichment, in maternal blood samples by fluorescence in situ hybridization (FISH) analysis for chromosomes X and Y (XY-FISH). In our blinded analysis of 40 maternal blood samples, we therefore examined all fetal cells without any enrichment. Initial examinations using conventional XY-FISH indicated a low specificity of 69.4%, which could be improved to 89.5% by the use of two different Y-chromosome-specific probes (YY-FISH) with only a slight concomitant decrease in sensitivity (52.4% vs 42.9%). On average, 12–20 male fetal cells/ml of maternal blood were identified by XY- and YY-FISH, respectively.

Trophoblasts Isolated from the Maternal Circulation: In Vitro Expansion and Potential Application in Non-invasive Prenatal Diagnosis

2005 ◽  
Vol 53 (3) ◽  
pp. 337-339 ◽  
Author(s):  
Esther Guetta ◽  
Liat Gutstein-Abo ◽  
Gad Barkai
Keyword(s):  
Prenatal Diagnosis ◽  
Maternal Blood ◽  
Male Fetus ◽  
Maternal Circulation ◽  
Fetal Cells ◽  
Non Invasive ◽  
In Vitro Expansion ◽  
Feasible Option

Prenatal diagnosis based on rare fetal cells in maternal blood is currently not a feasible option. An effort was made to improve cell yields by targeting trophoblast cells. After sorting, the HLA-G-positive cell fraction was analyzed directly or after culture. In situ hybridization technology was applied to prove fetal cell source in samples from women carrying a male fetus and to predict gender in samples without previous knowledge of fetal sex. In vitro culture led to a significant increase in fetal cells and accurate gender prediction in 93% of these samples. This approach might be useful for non-invasive prenatal diagnosis.

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

2014 ◽  
Vol 34 (6) ◽  
pp. 538-546 ◽  
Author(s):  
Ahmed Emad ◽  
Eric F. Bouchard ◽  
Josée Lamoureux ◽  
Annie Ouellet ◽  
Aparajita Dutta ◽  
...  
Keyword(s):  
Maternal Blood ◽  
Fetal Cells ◽  
Cellular Events ◽  
Microscope Slides ◽  
Automatic Scanning

Dual-Color PRINS for In Situ Detection of Fetal Cells in Maternal Blood

2006 ◽  
pp. 141-150
Author(s):  
Kada Krabchi ◽  
Macoura Gadji ◽  
Ju Yan ◽  
Régen Drouin
Keyword(s):  
Maternal Blood ◽  
Fetal Cells ◽  
Dual Color

scholarly journals Development of a Specific Monoclonal Antibody to Detect Male Cells Expressing the RPS4Y1 Protein

2021 ◽  
Vol 22 (4) ◽  
pp. 2001
Author(s):  
Silvia Spena ◽  
Chiara Cordiglieri ◽  
Isabella Garagiola ◽  
Flora Peyvandi
Keyword(s):  
Monoclonal Antibody ◽  
Prenatal Diagnosis ◽  
Maternal Blood ◽  
Limiting Factors ◽  
Inherited Disease ◽  
Specific Monoclonal Antibody ◽  
Native Form ◽  
Fetal Cells ◽  
Non Invasive ◽  
Reliable Isolation

Hemophilia is an X-linked recessive bleeding disorder. In pregnant women carrier of hemophilia, the fetal sex can be determined by non-invasive analysis of fetal DNA circulating in the maternal blood. However, in case of a male fetus, conventional invasive procedures are required for the diagnosis of hemophilia. Fetal cells, circulating in the maternal bloodstream, are an ideal target for a safe non-invasive prenatal diagnosis. Nevertheless, the small number of cells and the lack of specific fetal markers have been the most limiting factors for their isolation. We aimed to develop monoclonal antibodies (mAbs) against the ribosomal protein RPS4Y1 expressed in male cells. By Western blotting, immunoprecipitation and immunofluorescence analyses performed on cell lysates from male human hepatoma (HepG2) and female human embryonic kidney (HEK293) we developed and characterized a specific monoclonal antibody against the native form of the male RPS4Y1 protein that can distinguish male from female cells. The availability of the RPS4Y1-targeting monoclonal antibody should facilitate the development of novel methods for the reliable isolation of male fetal cells from the maternal blood and their future use for non-invasive prenatal diagnosis of X-linked inherited disease such as hemophilia.

A facile one pot route for the synthesis of imide tethered peptidomimetics

2016 ◽  
Vol 14 (2) ◽  
pp. 556-563 ◽  
Author(s):  
Veladi Panduranga ◽  
Girish Prabhu ◽  
Roopesh Kumar ◽  
Basavaprabhu Basavaprabhu ◽  
Vommina V. Sureshbabu
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Efficient Method ◽  
One Pot ◽  
Protected Amino Acid

A simple and efficient method for the synthesis of N,N’-orthogonally protected imide tethered peptidomimetics is presented. The imide peptidomimetics were synthesized by coupling the in situ generated selenocarboxylate of Nα-protected amino acids with Nα-protected amino acid azides in good yields.

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