Improved single-cell protocol for preimplantation genetic diagnosis of spinal muscular atrophy.

Abstract

OBJECTIVE:

To develop and validate a simple and reliable single-cell analysis protocol for the preimplantation genetic diagnosis (PGD) of spinal muscular atrophy (SMA).

DESIGN:

Molecular tests based on specific enzymatic digestion have already been described for SMA diagnosis. We modified the amplified DNA fragments so as to introduce a novel restriction site that provides an internal control for the completeness of the digestion.

SETTING:

The genetics and reproduction departments of two teaching hospitals.

PATIENT(S):

Six informed couples at risk of transmitting SMA.

INTERVENTION(S):

All patients underwent standard procedures associated with intracytoplasmic sperm injection.

MAIN OUTCOME MEASURE(S):

Improvement of SMA diagnostic efficiency and accuracy on single cell.

RESULT(S):

One hundred fifty lymphocytes were analyzed with our protocol. One hundred percent diagnostic accuracy was achieved from both homozygous normal and SMN1-deleted leukocytes. Successful molecular analysis was achieved for 36 of 42 biopsied embryos (86%). Twenty-five normal embryos were transferred, but no pregnancy was achieved.

CONCLUSION(S):

We developed an improved protocol for PGD of SMA that is simple, robust, and accurate; unfortunately, no pregnancies were achieved for any of the six patients who have undergone PGD in the program thus far.

• Authors: Philippe Burlet, Nelly Frydman, Nadine Gigarel, Jean Paul Bonnefont, Violaine Kerbrat, Gérard Tachdjian, René Frydman, Arnold Munnich, Julie Steffann, Pierre F Ray
• Journal: Fertility and sterility (Fertil Steril) Vol. 84 Issue 3 Pg. 734-9 (Sep 2005) ISSN: 1556-5653 [Electronic] United States
• PMID: 16169410 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Topics

• Base Sequence
• Embryo, Mammalian (cytology, physiology)
• Female
• Genetic Testing (methods, statistics & numerical data)
• Humans
• Lymphocytes (cytology, physiology)
• Molecular Sequence Data
• Muscular Atrophy, Spinal (diagnosis, genetics)
• Polymerase Chain Reaction (methods, statistics & numerical data)
• Pregnancy
• Preimplantation Diagnosis (methods, statistics & numerical data)