Non-invasive, prenatal, in-vitro method for detecting the normal healthy condition, the condition of a healthy carrier or the condition of a carrier inflicted with cystic fibrosis

摘要

Non-invasive, in vitroprenatal diagnosis of a normal healthy state; a healthy carrier or an affected carrier of cystic fibrosis (CF), comprises analysis of DNA-containing fetal cells (FC) isolated from a maternal sample. Non-invasive, in vitroprenatal diagnosis of a normal healthy state; a healthy carrier or an affected carrier of cystic fibrosis (CF) by analysis of DNA-containing fetal cells (FC) isolated from a maternal sample. The method comprises: (a) concentrating FC from the maternal sample; (b) analysis of retained cells and selecting those presumed to be of fetal origin; (c) confirming fetal origin of selected cells; (d) analyzing DNA of selected cells for alleles of the CFTR (= cystic fibrosis transmembrane conductance regulator) gene that contain the ~DF508, or other known, mutation, or for alleles of a locus carrying a polymorphism that is genotypically linked to an unidentified disease mutation. Step (d) comprises: (i) amplification of fetal DNA, using primers that can amplify a locus that may contain a known mutation or a locus as described above; (ii) identifying, on alleles that correspond to amplified DNA, presence or absence of a known mutation or locus; and (iii) comparing the fetal alleles with control samples to classify the CF status of the fetus. Independent claims are included for: (1) sixteen oligonucleotides (ON), sequences given in the specification, useful as primers for DNA amplification, also pairs of them and analogous sequences with at least 60, preferably 95, % identity; (2) combinations of pairs of external and internal primers (i.e. ON) for use, respectively, in first and second stage amplifications; and (3) a kit for the new process containing ON as primers, reagents for DNA amplification and/or instructions for use.