Maternal Alpha-Fetoprotein (AFP)

What sample is needed for maternal alpha-fetoprotein (AFP)?

1. Pregnant mother serum is required. It is stable for 24 hours at 2 to 8 °C.
2. Take 3 to 5 ml of blood in the disposable syringe.
   1. Keep the syringe for 15 to 30 minutes, then centrifuge for 2 to 4 minutes. This will yield a clear serum.

What precautions would you take for Maternal alpha-protein (AFP)?

• Keep serum at 2 to 8 °C if the test is performed within 24 hours; otherwise, freeze it at -20 °C.

What are the indications for Maternal alpha-protein (AFP)?

1. This is an effective screening marker for diagnosing body wall defects in the fetus, like neural tube defect,  spina bifida, or anencephaly.
2. AFP, on average, 25% to 30%, has a low value in Down’s syndrome.
3. This test is indicated in pregnant ladies who have the following findings:
   1. Female over the age of 35 years.
   2. If there is a family history of birth defects.
   3. If the lady has taken harmful medications during pregnancy.
   4. Female with a history of diabetes mellitus.

How will you define Alpha-fetoprotein (AFP)?

1. The fetal liver produces α1-globulin called alpha-fetoprotein (AFP).
2. AFP will become the dominant fetal protein in the first trimester, reaching its peak at 12 weeks gestation.
   1. During the first 10 weeks of fetal life, the major serum protein is not albumin; this is alpha-fetoprotein (AFP)
   2. AFP has the same sequence of amino acids as albumin. Suggesting the same gene ancestor.
3. Later on, it declines to 1% at birth.

What is the molecular structure and development of Alpha-fetoprotein (AFP)?

1. The fetal liver produces alpha-fetoprotein (AFP), which becomes the dominant fetal protein in the first trimester.
2. AFP peaks at 12 weeks of intrauterine fetal life and declines to 1% of the peak at birth.
3. AFP is a glycoprotein first produced by the yolk sac and then by the fetal liver.
   1. It reaches a peak at 10 to 13 weeks of gestation, later declining to <10 µg/L by the term.
   2. By the age of 2 years, it reaches the adult level (<5 µg/L)
4. AFP (α1- Fetoprotein) has 40% carbohydrates with a molecular mass of 70 kD. This is also called α1- Fetoprotein.
5. AFP is a glycoprotein that migrates more slowly on electrophoresis than albumin but is faster-moving than other globulins.
6. AFP is an oncofetal protein (glycoprotein) synthesized in the fetal liver and yolk sac.
7. AFP is very stable in serum, even at room temperature, for a week.
8. The AFP gene is located on the long arm (q) of chromosome 4 (q11 to q22). This is part of the family gene for albumin and vitamin D-binding protein.
9. Some of the fetal AFP enters the maternal serum (circulation).
10. AFP is the dominant fetal serum protein in the first trimester.
11. It is very low at the age of one year.

What are the important facts about Maternal Alpha-Fetoprotein (AFP)?

1. Maternal AFP rises progressively in the first and second trimesters.
2. Maternal serum peaks at the 13th week of gestation and then declines rapidly to <2% of the maximum level of 34 to 36 weeks of pregnancy.
3. The AFP is very high in the 8th week; then it dips at 11 weeks and again peaks at 13 weeks. Then, it falls in a long-linear fashion until 25 weeks.
4. Maternal AFP is 5 ng/mL at the 10th week of gestation, then increases by 15% per week.
5. It is 180 ng/mL at the 25th week, then declines slowly until term.
6. After birth, it drops to 2 ng/mL.

What is the Fetal AFP pattern?

1. In the early stages, AFP is produced in the yolk sac and is small in quantity. The fetal liver produces a major quantity and the yolk sac degenerates.
2. AFP has a high concentration in the fetal serum early in embryonic life.
3. The peak concentration is 3,000,000 ng/mL at 9 weeks of gestation. Then, concentration declines steadily to 20,000 ng/mL at term.
4. In infants, the AFP level declines and reaches adulthood by the 10th month of life.
5. By the age of one year, a more significant decrease occurred.

What are the Functions of the AFP?

1. Amniotic AFP is more accurate than maternal serum AFP in diagnosing neural tube defects in early gestation (around 14 weeks).
   1. Neural tube defects vary from small myelomeningocele to anencephaly.
2. Other fetal body wall defects are:
   1. Omphalocele.
   2. Gastroschisis.

Maternal alpha-protein (AFP) and Neural tube defect

3. Before 14 weeks, AFP helps to diagnose neural tube defects.
   1. Normal AFP <2.5 Multiples of the median (MoM).
   2. For the neural tube, the defect is >2.5 MoM.
4. If there is a body wall defect in the fetus, AFP leaks out into amniotic fluid and is picked up by the maternal circulation.
5. If the AFP level is increased, repeated AFP, amniotic fluid AFP, and ultrasound are recommended for further evaluation.
   1. The serum AFP level is 100 times that of amniotic fluid.
6. AFP is used to diagnose neural tube defects.
7. Raised AFP level may indicate multiple pregnancies, fetal distress, congenital fetal abnormalities, or intrauterine death.
8. A Low AFP level may diagnose Down’s syndrome (trisomy 21).
9. AFP is used as a tumor marker.

How would you interpret Maternal serum screening of the alpha-fetoprotein (AFP)?

1. There is the possibility of 3% birth defects in newborns.
2. Early detection of the fetus abnormality is the goal of maternal serum AFP estimation.
3. In most cases, AFP estimation is recommended during 16 to 18 weeks of gestation, and the specimen is collected during 14 to 20 weeks of gestation.
4. If it is delayed, then deciding to terminate the pregnancy is difficult, like elective termination of pregnancy.
5. In the case of neural tube defect, there is increased fetal amniotic fluid and mother serum AFP.
   1. Maternal serum AFP helped diagnose open neural tube defects.
   2. 90% of newborns with neural tube defects have no known risk factors.
6. In Down’s syndrome, the maternal serum AFP is low.
7. Tripple screening tests done in the second trimester to find potential birth defects include:
   1. Maternal Serum AFP.
   2. Maternal HCG.
   3. Maternal Estriol level.

What are the normal values of Alpha-fetoprotein (AFP)?

Source 2

• Adult = <40 ng/mL (<40 mcg/L)
• Child (<1 year) = <30 ng/mL

Another source

• 25 ng/mL (25 µg/L).
• At 15 to 18 weeks of gestation = 10 to 150 ng/mL (10 to 150 µg/L).

What are the causes of the raised Alpha-fetoprotein (AFP) level?

1. Maternal serum level >2 times the median level will be seen in:
   1. Multiple gestations (multiple pregnancies).
   2. Fetal death.
   3. Malformations, e.g., open neural tube defects like anencephaly, open spinal Bifida, encephalocele, and myelocele.
   4. 80% of the cases are diagnosed by AFP level (hydrocephaly and microcephaly).
2. Maternal serum and amniotic fluid Increased AFP concentration:
   1. Open neural tube defects like anencephaly, spina bifida, omphalocele, esophageal or duodenal atresia.
   2. Threatened abortion.
   3. Fetal distress.
   4. Intrauterine death of the fetus.
   5. Fetal congenital abnormalities.
   6. Abdominal wall defects like gastroschisis.
3. Other conditions for raised Alpha-fetoprotein (AFP) are:
   1. Renal abnormalities
   2. Cystic hygroma.
   3. Hydrops fetalis.
   4. Turner syndrome.
   5. Bowel obstruction.
   6. Twins.
   7. Feto-maternal hemorrhage.
   8. Sacrococcygeal Teratoma.
   9. Esophageal or duodenal atresia.
   10. Renal disorders like polycystic kidney, renal agenesis, and urethral obstruction.
   11. Tetralogy of Fallot.
   12. Oligohydramnios.
   13. Turner syndrome.
   14. Placental causes include thrombosis, infarction, large placenta, and cystic changes.
   15. Maternal causes like tumor-producing AFP and hepatitis.

What are the causes of decreased Maternal Alpha-Fetoprotein (AFP)?

1. Down syndrome (Trisomy 21).
2. Long-standing death of the fetus.
3. Molar pregnancy (Hydatidiform mole).
4. Choriocarcinoma.
5. Spontaneous abortion.
6. Overestimated gestational age.
7. Pseudopregnancy.
8. A low level of AFP with an abnormal value of HCG and estriol (Triple screening) is indicative of :
   1. Trisomy 21 (Down’s syndrome).
   2. Trisomy 18 (Edwards syndrome)
   3. Or other chromosomal abnormalities.
   4. Women with diabetes mellitus have 20% to 40% lower values than nondiabetic women.

What are the causes of increased maternal Alpha-fetoprotein (AFP)?

1. It is advised to do amniotic fluid AFP.
2. Multiple pregnancies.
3. Open neural tube defects like:
   1. Open spina bifida.
   2. Encephalocele.
   3. Myelocele.
   4. Anencephaly.
   5. Sacrococcygeal teratoma.
4. Turner syndrome.
5. Omphalocele.
6. Gastroschisis.
7. Hydrops fetalis.
8. Intrauterine death.
9. Feto-maternal hemorrhage.
10. Esophageal or duodenal atresia.
11. Tetralogy of Fallot.
12. Oligohydramnios.
13. Cystic hygroma.
14. Placental causes are:
    1. Thrombosis.
    2. Infarction.
    3. Cystic changes.
    4. Infections.
    5. Very large placenta.
15. Rarely benign hereditary familial raised level of AFP.
16. Maternal causes:
    1. Hepatitis.
    2. Malignancies producing AFP.

Markers to detect fetal abnormalities:

Questions and answers: