Since March 2004, a new law regulates IVF techniques in Italy, imposing very strict conditions (Benagiano and Gianaroli, 2004). This law limits to three the number of oocytes that can be fertilized during each IVF treatment and obligates the simultaneous transfer of all three possible embryos. Oocyte cryopreservation is allowed while freezing of spare embryos is prohibited. Moreover, the use of IVF techniques is restricted to sterile couples, thus precluding the access to assisted reproductive techniques to fertile couples who are at risk for transmitting severe genetic diseases to their offspring.
According to the text of the new regulation, technically PGD would be not illegal, because “clinical and experimental research on embryos is permitted only for therapeutic and diagnostic purposes”. However, the obligation for transfer of all embryos generated after oocytes’ fertilization, including those resulting affected by the genetic disease, makes PGD unfeasible. Afterwards, the Ministry of Health Guidelines has clarified that PGD on embryos is forbidden for any purposes.
Therefore, the only option for couples at high genetic risk for prevention of genetic diseases is 1PB testing (so called Preconception genetic diagnosis) before oocyte fertilization, provided that they are also infertile.
In preconception genetic diagnosis, ICSI can be performed only after genetic diagnosis of oocytes. Moreover, results of genetic testing must be achieved within a reasonable time to prevent in-vitro ageing of the oocytes. In fact, there is only a very narrow window of time available for preconception genetic diagnosis, but if the 1PB biopsy is performed soon after oocyte collection (Magli et al., 2006) and follows a rapid diagnostic protocol, oocyte insemination could be carried out according to the results of the genetic analysis.
We have overcome to the time restriction problem by developing a rapid protocol for diagnosis of single gene mutations of maternal origin, capable of producing results within just 4 hours, making it realistic to fertilize the oocytes predicted to be free of mutation within a timeframe compatible with a late ICSI (?6 h after oocytes collection).