Fertility and sterility · 2005
Improved single-cell protocol for preimplantation genetic diagnosis of spinal muscular atrophy.
СтатьяПоддержка исследования: вне правительства СШАНазвание и аннотация: Перевод пока не подготовлен
Обработка статьи · этапы, ошибки и управление
Загрузка состояния…
Ошибки и ограничения
История · было → стало
Состояние обновляется каждые 5 секунд, пока блок открыт. Процент показывается только там, где известен объём работы. Пауза останавливает продолжение этапа; текущий запрос к источнику может завершиться.
Burlet Philippe, Frydman Nelly, Gigarel Nadine, Bonnefont Jean Paul, Kerbrat Violaine, Tachdjian Gérard, Frydman René, Munnich Arnold, Steffann Julie, Ray Pierre F
Аннотация
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.
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.