Clinical application of multiple displacement amplification in preimplantation genetic diagnosis.
Multiple displacement amplification (MDA) is a technique used in the amplification of very small amounts of DNA. MDA is reported to yield large quantities of high-quality DNA. The applicability of MDA to single cells was recently demonstrated as a potential technique for preimplantation genetic diagnosis (PGD). This paper shows the first clinical application of MDA in PGD. Two cycles of PGD were performed in two diseases, resulting in two pregnancies. All the diagnoses given on blastomeres were confirmed on the non-transferred whole embryos. The blastomere diagnosis was coupled with short tandem repeat (STR) analysis (16 loci) in all cycles. Allelic drop-out (ADO) assessment and amplification efficiency were evaluated on 40 single lymphocytes derived from parents of each disease. ADO and amplification failure were 10.3 and 2.2% for beta-thalassaemia and 17.9 and 2.2% for cystic fibrosis respectively. HLA matching for A, B and DR was performed successfully on single cell for the beta-thalassaemia family using similar methods to genomic DNA. The PGD protocol used in all diseases consists of MDA amplification, followed by a standard polymerase chain reaction protocol. Although HLA matching was not applied to embryos, its feasibility was shown on single cell DNA amplified by MDA. Altogether, these data show the simplicity and reliability of performing PGD in combination with HLA matching and STR analysis using MDA.
Hellani
Ali
A
King Faisal Specialist Hospital and Research Centre, Riyadh, 11211 KSA. hellani@kfshrc.edu.sa
Coskun
Serdar
S
Tbakhi
Abdelghhani
A
Al-Hassan
Saad
S
D003550:Cystic Fibrosis / Q000175:diagnosis / Q000235:genetics / Q000276:immunology; D005260:Female; D006239:Haplotypes; D006650:Histocompatibility Testing; D006801:Humans; D008297:Male; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D011247:Pregnancy; D019836:Preimplantation Diagnosis / Q000379:methods*; D020080:Tandem Repeat Sequences; D017086:beta-Thalassemia / Q000175:diagnosis / Q000235:genetics / Q000276:immunology