Cystic fibrosis - prenatal diagnosis

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Cystic Fibrosis

Identification of disease Cystic fibrosis is an autosomal recessive genetic disorder that is caused by mutations in the cystic fibrosis transmembrane regulator (CFTR) gene on chromosome 7 which codes for the protein that controls ion transfer across cell membranes. This disease is more common in Caucasian people - About 5 percent of white people carry one defective gene responsible for cystic fibrosis, but the trait is recessive, and the disease develops only if a person has two defective genes. People with only one defective gene have no noticeable symptoms. Since this is not a sex- linked disorder, the carriers of this disease are both female & male, provided that they are heterozygous

Symptoms Cystic fibrosis (CF) is caused by changes in a protein that controls the transfer of chloride and sodium ions (salts) across cell membranes. Disruption of salt transfer results in abnormal gland secretions and dehydration due to increased loss of salt and water during sweating. CF affects almost all of the glands in the body that secrete fluid, resulting in a variety of symptoms. Secretions may be thick and cause blockage in the pancreas, intestines and lungs. Mucus blockage also provides places for bacteria to multiply, increasing the probability of infection. CF children show poor digestion, dehydration, coughing and vomiting. As the disease progresses, teenagers show slowed growth, delayed puberty and reduced physical endurance. Adults show more serious complications such as collapsed lung, heart failure, infertility and frequent infections that eventually lead to death.

Prenatal Diagnosis of the disease-

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a) Purpose & Usefulness Improved techniques of prenatal diagnosis give parents ( Especially those who have a known family case) the opportunity to find out whether or not the fetus has CF. IF the result is positive, then this Prenatal diagnosis of CF would allow the parents time to personally make a decision regarding the termination of the pregnancy. Even if the parents refuse to terminate the pregnancy, the knowledge that their child will be suffering from Cf will be of great help It would help them to prepare and cope with the implications of the situation which they will be facing when the child is born.

b) Types of diagnosis Diagnosis can be made by chorionic villus sampling in the tenth to twelfth weeks of pregnancy. A later test (amniocentesis) is available at 16 weeks.

c) Techniques/procedures -

CVS Chorionic villus sampling (CVS) provides the ability to obtain fetal tissue from the developing trophoblast for diagnostic studies. The chorion frondosum, which contains the most mitotically active cells, is the area that is sampled. CVS can be accomplished using either a transcervical or transabdominal approach. Prior to the procedure, an ultrasound is performed to assess fetal viability, gestational age and placental position. Transcervical CVS involves passing a polyethylene catheter with a malleable obturator through the cervix to the thickest part of the placenta using ultrasound guidance. Placental trophoblast is then aspirated through the catheter into a syringe that contains tissue culture medium. In Transabdominal CVS, a needle is placed through the long axis of the placenta under ultrasound guidance. The stylet is withdrawn from the needle, a syringe containing tissue culture medium is attached to the hub of the needle and suction is applied as the needle is moved up and down through the placenta until an adequate amount of tissue is obtained.

Amniocentesis With the use of an ultrasound scanning techniques the position of the fetus and the location of the placenta are determined. The skin on the mother's abdomen is aseptically prepared and a local anesthetic is usually injected. A needle attached to a syringe is introduced into a part of the uterus where there is the least chance of perforating the placenta or scratching the fetus. Between 0 & 5 ml of amniotic fluid is aspirated.

The Tissue obtained from CVS and the amniotic fluid obtained from amniocentesis could then be used by direct DNA testing to detect the CFTR gene mutation. Direct Dna testing is carried out using the following steps-

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