The overall goal of this project is to develop a rapid, non-invasive and sensitive system for pre-natal testing for fragile X syndrome (FXS). Asuragen will combine our unique and well-characterized technologies for accurate detection of repeat expansions with Savran Technologies' novel immunomagnetic Micro-Aperture Chip (IMAC) system to enable prenatal testing of circulating fetal trophoblasts (cTB) isolated from maternal whole blood. The combination of these systems will accomplish fetal cell selection using a novels system for antibody- coupled magnetic fractionations that generates the substrate for highly sensitive and accurate sizing of FMR1 CGG repeats using Asuragen's AmplideX reagents and workflows. The American College of Obstetricians and Gynecologists clinical guidelines recommend prenatal testing for FXS. Women who carry an FMR1 premutation or full mutation should be offered prenatal diagnosis because these women are at risk for having children with FXS. Collectively, more than one million women in the US may be carriers, and those who are pregnant could benefit from noninvasive prenatal testing. The system developed under proposal would enable prenatal screening using maternal blood samples, reducing the risks associated with aminocentesis and chorionic villus sampling (risk of miscarriage of up to 1 in 100) and permitting testing to be performed as early as 5 weeks of gestation. The specific aims are: Aim 1: Establish the feasibility of the IMAC system for immunomagnetic separation for trophoblast cells from cell lines spiked into whole blood for characterization by AmplideX PCR reagents in a workflow that includes Whole Gene Amplification (WGA). Aim 2: Demonstrate characterization of IMAC fractionated cells from 11 cell lines with well-characterized genotypes across the full range of FMR1 allele-specific sizing representing normal, permutation, and expanded alleles. Aim 3: Optimize performance of the IMAC/WGA/AmplideX workflow and demonstrate feasibility of prenatal testing by characterization of cTB from residual human blood specimens.
Public Health Relevance Statement: Project Narrative This project aims to advance pre-natal testing of women at risk of having a child with fragile X syndrome by combining innovative reagents for repetitive sequence detection with a novel system for cell enrichment. The combined system will enable accurate screening using maternal blood samples in order to reduce risk of miscarriage and provide screening much earlier in the pregnancy.
Project Terms: Alleles; American College of Obstetricians and Gynecologists; Amniocentesis; Antibodies; Binding; Biological Assay; Blood Cells; Blood Circulation; Blood specimen; Categories; Cell Line; Cell Separation; Cells; CGG repeat; Chemistry; Child; Chorionic Villi Sampling; Clinical; Coupled; Detection; Diagnosis; Disease; fetal; fetus cell; FMR1; FMR1 Premutation; Fractionation; Fragile X Syndrome; Gene Amplification; Genotype; Goals; Guidelines; high risk; Human; Immunomagnetic Separation; innovation; Liquid substance; lymphoblastoid cell line; magnetic field; Magnetism; Modeling; Mutation; new technology; novel; particle; Performance; performance tests; Pregnancy; pregnant; Prenatal Diagnosis; prenatal testing; Procedures; Reagent; Recovery; Repetitive Sequence; Research; Residual state; Risk; Sampling; screening; Spontaneous abortion; Surface; Surface Antigens; System; Technology; Testing; Time; trend; trophoblast; Universities; Whole Blood; Woman
