In Vitro Fertilisation : In order to achieve pregnancy, the sperm has to unite with the egg, which is released from the ovary. Normally this union is called fertilization, it occurs with in the fallopian tube. However, in IVF the union occurs in the laboratory in which egg and sperm have been collected and cultured together to form embryos and then embryos are transferred to the uterus on day 3 to continue growth. This process is called In vitro Fertilisation.
The growth and development of the eggs is closely monitored by repeated ultrasound studies and ovulation itself is triggered by an injection (Human Chorionic Gonadotropin), after which the egg retrieval is scheduled.
Procedure: five major steps in the IVF and embryo transfer sequence:
(Collection of Eggs): Eggs are gently removed from ovaries using the needle. This is called "follicular aspiration."They are placed with the sperm in incubators to allow fertilization to take place.
Obtaining the sperms: The semen from man is obtained by masturbation in a sterile container and handed over to embryologist for the processing of semen, for separation of sperms from the semen plasma, other debris and immotile cells.The processed active motile sperms are selected and used for IVF /ICSI procedures. For patients /couples having recurrent failures in implantation and miscarriages it is mandatory to undergo DNA fragmentation test before undergoing IVF/ICSI procedures.
DNA Fragmentation Test: We can assess cell death by detecting DNA fragments, using agarose gel electrophoresis. One of the classic features of apoptosis is the cleavage of the genomic DNA into oligonucleosomal fragments.It is represented by multiples of 180-200bp.
This test may help to identify men who are at risk of failing to initiate a pregnancy.Information about sperm DNA integrity may help in the clinical diagnosis, management and treatment of male infertility and may be of prognostic value in assessing outcome of assisted conception treatment.
Fertilisation: The eggs are placed with the sperm in incubators to allow fertilization to take place. Normally in IVF one egg is mixed with 100,000 sperms and one of the sperms fertilizes the egg on its own. The eggs are examined carefully at intervals to ensure that fertilization and cell division have taken place. The fertilized eggs are now called embryos. Embryos are usually placed in the women’s uterus 2 or 3 days after egg retrieval. The patient is informed to consult the doctor for Serum BHCG test after 14 days of medication or, can do it in any lab nearby and can inform the obtained result value to confirm the pregnancy.