Reliability of prenatal diagnosis of genetic diseases by analysis of amplified trophoblast DNA.

Abstract

Dot blot analysis on enzymatical] trophoblast DNA with allele specific tide probes is currently used for diagnosis of single gene disorders cha the molecular level, such as the ,B th phenylketonuria, sickle celi anaemia, trypsin deficiency. A potential problem of this procedure is the co-amplificatioi sequences, which may obscure the dia fetus. To address this question, we prenatal diagnosis of ,B thalassaemia in at risk by dot blot analysis on enzymatic; DNA with 32p or horseradish peroxi' allele specific oligonucleotide probes. the diagnosis obtained by this procedw nucleotide hybridisation on electri separated non-amplified trophoblast ments. We detected no co-amplifi sequences, even with a faint signal, in of trophoblast DNA from those fetus as normal or homozygotes, nor in tho as heterozygotes, who were born to par different mutations and had inherited mutation. These results indicate that, dissection of trophoblast tissue fro decidua is carried out, amplification villi DNA is not associated with aml maternal DNA sequences. We may tl that dot blot analysis of trophoblast D reliable procedure for prenatal diagno4 Three hundred pregnant women of Italian descent requesting prenatal diagnosis because they were at risk for thalassaemia major were included in this mber 1989. study.

From Prenatal to Preimplantation Genetic Diagnosis of β-Thalassemia. Prevention Model in 8748 Cases: 40 Years of Single Center Experience

“…After 1985 we introduced PCR by RDB, allelic-specific oligonucleotide primers by ARMS. So far, we have not had any misdiagnosis by TA-CVS using also only a few amplification cycles and two different overlapping amplified DNA fragments in duplicate analyses [ 54 ].…”