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Feasibility Study for a Microchip‐Based Approach for Noninvasive Prenatal Diagnosis of Genetic Diseases
Author(s) -
CREMONESI L,
GALBIATI S,
FOGLIENI B,
SMID M,
GAMBINI D,
FERRARI A,
VIORA E,
CAMPOGRANDE M,
PAGLIANO M,
TRAVI M,
PIGA A,
RESTAGNO G,
FERRARI M
Publication year - 2004
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1196/annals.1318.017
Subject(s) - fetus , prenatal diagnosis , cell free fetal dna , polymerase chain reaction , allele , genetic testing , andrology , chorionic villus sampling , biology , dna , gestational age , digital polymerase chain reaction , pregnancy , medicine , gene , obstetrics , genetics
A bstract : Fetal DNA in maternal plasma may represent a source of genetic material for prenatal noninvasive diagnosis of genetic diseases. We evaluated a cohort of physiological pregnancies to determine if fetal DNA can be retrieved at any gestational week in sufficient quantity to be analyzed with advanced mutation detection technologies. We performed fetal DNA quantification by real‐time polymerase chain reaction (PCR) on the SRY gene in 356 women sampled from 6 to 40 gestational weeks. Fetal DNA was retrieved at any week. All female fetuses were correctly identified. In 5 of 188 (2.6%) male‐bearing pregnancies, no amplification was obtained. For noninvasive testing, complete clearance of fetal DNA after delivery is mandatory. Long‐term persistence was not detected in women with previous sons or abortions. These findings confirm that maternal plasma may represent the optimal source of fetal genetic material. For noninvasive diagnosis of genetic diseases, we evaluated microchip technology. The detection limit for a minority allele determined by diluting a mutated DNA into a wild‐type plasma sample was 5 genome equivalents, indicating that the test might be applied to the identification of paternally inherited fetal alleles in maternal plasma. The addition of peptide nucleic acids (PNAs) to either the PCR reaction or the chip hybridization mixture allowed approximately 50% inhibition of wild‐type allele signals.