Also Known As
AFP Maternal
Maternal Serum AFP
MSAFP
msAFP
Triple Screen
Triple Test
Quad Screen
Quadruple Marker Test
4-marker Screen
Multiple Marker Test
Formal Name
Maternal Serum Screen
This article was last reviewed on
This article waslast modified on
January 15, 2018.
At a Glance
Why Get Tested?

For pregnant women, to assess the risk of having a baby with certain chromosomal disorders, such as Down syndrome (trisomy 21) or Edwards syndrome (trisomy 18), or open neural tube defects, such as spina bifida

When To Get Tested?

Usually between the 15th and 20th weeks of pregnancy

Sample Required?

A blood sample drawn from a vein in your arm

Test Preparation Needed?

None

You may be able to find your test results on your laboratory's website or patient portal. However, you are currently at Lab Tests Online. You may have been directed here by your lab's website in order to provide you with background information about the test(s) you had performed. You will need to return to your lab's website or portal, or contact your healthcare practitioner in order to obtain your test results.

Lab Tests Online is an award-winning patient education website offering information on laboratory tests. The content on the site, which has been reviewed by laboratory scientists and other medical professionals, provides general explanations of what results might mean for each test listed on the site, such as what a high or low value might suggest to your healthcare practitioner about your health or medical condition.

The reference ranges for your tests can be found on your laboratory report. They are typically found to the right of your results.

If you do not have your lab report, consult your healthcare provider or the laboratory that performed the test(s) to obtain the reference range.

Laboratory test results are not meaningful by themselves. Their meaning comes from comparison to reference ranges. Reference ranges are the values expected for a healthy person. They are sometimes called "normal" values. By comparing your test results with reference values, you and your healthcare provider can see if any of your test results fall outside the range of expected values. Values that are outside expected ranges can provide clues to help identify possible conditions or diseases.

While accuracy of laboratory testing has significantly evolved over the past few decades, some lab-to-lab variability can occur due to differences in testing equipment, chemical reagents, and techniques. This is a reason why so few reference ranges are provided on this site. It is important to know that you must use the range supplied by the laboratory that performed your test to evaluate whether your results are "within normal limits."

For more information, please read the article Reference Ranges and What They Mean.

What is being tested?

Maternal serum screening, or the multiple marker test, measures several substances in the blood of a pregnant woman in the second trimester of pregnancy. The results can be used to determine the risk that the baby has a chromosome disorder or neural tube defect such as spina bifida.

These substances include the following, listed below. When the first three substances are measured together, it is called a triple screen. When the fourth substance, inhibin A, is added, it is called a quad screen.

  • Alpha-fetoprotein (AFP) is a protein produced by fetal tissue. During development, AFP levels in fetal blood and amniotic fluid rise until about 12 weeks, and then levels gradually fall until birth. Some AFP crosses the placenta into the mother's blood. With certain problems affecting a baby, an increased amount of AFP leaks into amniotic fluid and then into the mother's blood. As such, an AFP test is used to screen for neural tube defects such as spina bifida. It may be performed as part of the triple or quad screen. It may be performed by itself when risk for chromosome disorders (Down syndrome (trisomy 21) and Edwards syndrome (trisomy 18)) have already been assessed using first trimester Down syndrome screening or cell-free fetal DNA testing. According to the American College of Obstetricians and Gynecologists (ACOG), the AFP test detects neural tube defects in 85% of the cases.
  • Human chorionic gonadotropin (hCG) is a hormone produced by the placenta. Levels in maternal blood rise for the first trimester of pregnancy and then decrease during the remainder of the pregnancy. In pregnancies where the fetus is carrying the chromosomal defect that results in Down syndrome, hCG tends to be high whereas in pregnancies where the fetus has the chromosomal defect that results in Edwards syndrome, hCG tends to be low.
  • Unconjugated estriol (uE3) is a form of estrogen that is produced by the fetus through metabolism. This process involves the liver, adrenals, and the placenta. Some of the unconjugated estriol crosses the placenta and can be measured in the mother's blood. Levels rise around the 8th week and continue to increase until shortly before delivery. In pregnancies where the fetus has Down syndrome or Edwards syndrome, uE3 tends to be low.
  • Inhibin A is a hormone produced by the placenta. Inhibin is a dimer (has two parts) and is sometimes referred to as DIA or dimeric inhibin A. Levels in maternal blood decrease slightly from 14 to 17 weeks gestation and then rise again. Levels tend to be higher in pregnancies where the fetus has Down syndrome.

Including the fourth marker, inhibin A, increases both the sensitivity and specificity of the screen for Down syndrome. According to ACOG, the triple screen detects Down syndrome in 69% of the cases while the quad screen detects it in 81% of the cases.

If the results of maternal serum screening are cause for concern, diagnostic tests such as amniocentesis or chorionic villus sampling (CVS) may be recommended.

How is the sample collected for testing?

Blood is drawn from a vein in the woman's arm.

Is any test preparation needed to ensure the quality of the sample?

No test preparation is needed.

Accordion Title
Common Questions
  • How is it used?

    Maternal serum screening is a group of tests used in the second trimester of pregnancy to help evaluate a woman's risk of carrying a baby with chromosome disorders, including Down syndrome (trisomy 21) or Edwards syndrome (trisomy 18), or neural tube defects such as spina bifida or a condition called anencephaly.

    The tests are often combined into a triple or quad screen because their value lies in their use together. A mathematical calculation involving the levels of these substances (AFP, hCG, unconjugated estriol, and, sometimes, inhibin A) as well as considerations of maternal age, family history, weight, race, and diabetic status is used to determine a numeric risk for abnormalities in the fetus.

    The second trimester maternal serum screen is one of the options that may be offered for prenatal screening for fetal abnormalities. Others options include the first trimester screen and cell-free fetal DNA (cffDNA) testing.

    An AFP test may be performed by itself and not as part of a triple or quad screen, especially when first trimester screening or cffDNA testing has already been used to assess the risk for a chromosomal disorder. The AFP is used to help determine the risk of neural tube defects.

    The screening approach a woman may choose depends on what technology is available and when she first seeks prenatal care. For more information about how maternal serum screening fits in with other screening options, see below.

  • When is it ordered?

    Your healthcare practitioner may offer you the test between the 15th and 20th weeks of pregnancy.

  • What does the test result mean?

    A genetic counselor or healthcare practitioner who can explain the meaning of the results and offer choices about follow up should interpret the screening results.

    It is important to remember that positive screening tests are not diagnostic of a fetal abnormality. While they indicate an increased risk, only a small number of women who have positive maternal serum screening results have babies who actually have a neural tube defect or chromosomal abnormality.

    In pregnancies where the fetus is carrying the chromosomal defect that results in Down syndrome (trisomy 21), the levels of AFP and unconjugated estriol tend to be low and hCG and inhibin A levels high.

    In pregnancies where the fetus has Edwards syndrome (trisomy 18), unconjugated estriol and hCG levels are low and AFP levels can be variable.

    A baby with an open neural tube defect has an opening in its spine or head that allows higher-than-usual amounts of AFP to pass into the mother's blood. The other markers are not used in the evaluation of risk for carrying a fetus with a neural tube defect.

    Second Trimester Maternal Serum Screening Analyte Patterns
    Increased risk for: hCG uE3 AFP Inhibin A
    Open spina bifida Normal Normal High Not applicable
    Anencephaly Low Low High Not applicable
    Down syndrome High Low Low High
    Edwards syndrome Low Low Variable Not applicable

    If a screen is positive, tests that are more definitive are needed to determine and confirm a diagnosis. These include high-resolution ultrasound and perhaps amniocentesis followed by chromosome analysis. These follow-up tests are used to help women and their healthcare practitioners make decisions about managing their pregnancies.

    Screening will not detect all cases of fetal abnormalities.

  • Is there anything else I should know?

    The test result is very dependent on accurate determination of the gestational age of the fetus. If the gestational age of the fetus has not been accurately determined, the results may be either falsely high or low.

    In multiple gestation pregnancies, calculation of the risk of Down syndrome or Edwards syndrome is difficult. For twin pregnancies, a "pseudorisk" can be calculated comparing results to normal results in other twin pregnancies.

    Evaluation of the risk of open neural tube defects in twin pregnancies can be determined, although it is not as effective as in singleton pregnancies. For women further along in their pregnancies, risk cannot be calculated from these tests.

    If you have had a first trimester Down syndrome screen, then second trimester maternal serum screening is typically not performed because the risks for Down syndrome and Edwards syndrome have already been assessed. However, if you and your healthcare practitioner wish to use the results of both first trimester and second trimester screening to assess the risk of chromosome abnormalities, then integrated or sequential screening may be employed. For more on these, see below.

  • What is Down syndrome?

    Down syndrome (DS) is a chromosomal abnormality that is also called trisomy 21 because those with DS have an extra copy of part or all of chromosome 21. About 1 in 700 babies are born with Down syndrome each year in the U.S. The condition causes mild to moderate mental retardation and developmental problems and can be associated with congenital heart defects, respiratory and hearing problems, leukemia, and thyroid disorders. Many of the complications of Down syndrome can be treated and the lifespan of those affected has greatly increased in recent years. The risk of having a child with Down syndrome increases with the age of the mother. Although the risk of having an affected baby is significantly greater in those older than 35, the majority of Down syndrome babies (about 80%) are born to those under 35 because this age group has the greatest number of children. For this reason, the American College of Obstetricians and Gynecologists has recommended that all pregnant women be offered a screening test for Down syndrome.

  • What is Edwards syndrome?

    Edwards syndrome (trisomy 18) is a condition in which there are three copies of chromosome 18. As with Down syndrome, the risk of carrying a fetus with Edwards syndrome increases with maternal age. Edwards syndrome is associated with multiple abnormalities and is usually fatal, with live-born infants rarely living beyond one year of age. The frequency of this abnormality is much less than that of Down syndrome, occurring in only 1 in 6,000 live births. For more information, see the Trisomy 18 Foundation web site.

  • What is a neural tube defect?

    Neural tube defects (NTDs) are serious birth defects in which the brain, spinal cord, or their coverings do not develop completely. They arise early in pregnancy, affecting fetal development, and can cause life-long complications of varying severity for the child. There are several types of neural tube defects, the most common of which are:

    • Spina bifida: the most common neural tube defect; occurs when the neural tube does not close completely somewhere along the spine
    • Anencephaly: occurs when the neural tube fails to close properly at the head; results in incomplete development of a large portion of the brain and the skull
  • What can I do to help prevent formation of a neural tube defect?

    The U.S. Public Health Service and the Centers for Disease Control and Prevention recommend that all women of childbearing age take 0.4 mg (400 micrograms) of folic acid daily. They recommend this to all women between the ages of 15 and 45 because many pregnancies are unplanned and neural tube defects occur very early in a pregnancy, before a woman may realize that she is pregnant. The recommended dose may be higher for women considered at high risk for having a baby with a neural tube defect, such as those who have already had an NTD-affected pregnancy. Folic acid can be found in fortified foods, supplements, and from a healthy, varied diet. For more information, see the CDC's web page Facts About Folic Acid.

  • Are there other non-invasive screening tests for these disorders?

    A newer test called cell-free fetal DNA (cffDNA) only requires a blood sample from the pregnant woman and can be used to screen for certain fetal chromosomal abnormalities, including Down syndrome, Edwards syndrome, and Patau syndrome (trisomy 13). It can be performed as early as the tenth week of pregnancy. It does not, however, screen for neural tube defects like spina bifida. Invasive diagnostic tests, such as chorionic villus sampling (CVS) and amniocentesis, are still needed to confirm the results of cffDNA. Since cffDNA screening is a relatively new test, some insurance companies may not cover it and it may not be available everywhere. For more information, see the article on Cell-Free Fetal DNA.

  • I am pregnant. Will I have both first and second trimester screening performed?

    There are several approaches to screening depending on what technology is available where you are and when you first visit your healthcare practitioner for prenatal care. Options include:

    First trimester screening for chromosome abnormalities using the combined tests for pregnancy-associated plasma protein A (PAPP-A), hCG and nuchal translucency ultrasound, followed by a blood test for maternal AFP and/or a fetal ultrasound in the second trimester to check for neural tube defects

    • Second trimester screening (triple or quad screen) to assess risk for chromosome abnormalities and neural tube defects; this may be the only testing done, especially in the case of a woman who does not visit her healthcare practitioner until the second trimester.

    Cell-free fetal DNA testing in the first or second trimester with a blood test for maternal AFP and/or a fetal ultrasound in the second trimester to check for neural tube defects

    Typically, if first trimester screening for chromosome disorders has been performed, then screening in the second trimester is not performed because the risks for Down syndrome and Edwards syndrome have already been assessed. However, if you and your healthcare practitioner wish to use the results of both first trimester Down syndrome screen combined tests and the second trimester maternal serum screen to assess the risk of chromosome abnormalities, then one of the following approaches is used:

    • Integrated screening – involves performing both first and second trimester testing and not releasing the results until all testing is completed

    • Sequential screening – involves performing a first trimester screen; if the screen indicates increased risk for Down syndrome or Edwards syndrome, a diagnostic procedure such as chorionic villus sampling is offered. If the screen does not indicate increased risk, then a triple or quad screen in the second trimester is offered and results from both the first and second trimester screens are used in the final risk assessment.

View Sources

Sources Used in Current Review

(2014 April). American College of Obstetricians and Gynecologists. Screening tests for birth defects, FAQ.  Available online at http://www.acog.org/Patients/FAQs/Screening-Tests-for-Birth-Defects. Accessed 12/26/2016.

(2015 September). American College of Obstetricians and Gynecologists. Committee Opinion, Number 640. Available online at http://www.acog.org/Resources-And-Publications/Committee-Opinions/Committee-on-Genetics/Cell-free-DNA-Screening-for-Fetal-Aneuploidy. Accessed 12/26/2016.

Mayo Clinic staff. (2015 October 21). Quad screen. Mayo Clinic. Available online at http://www.mayoclinic.com/health/quad-screen/MY00127. Accessed 12/26/2016.

(Reviewed 2016 April 21). Centers for Disease Control and Prevention. Folic acid, recommendations. Available online at https://www.cdc.gov/ncbddd/folicacid/recommendations.html. Accessed 12/26/2016.

(2016 May). American College of Obstetricians and Gynecologists. Practice Bulletin Number 163: Screening for fetal aneuploidy. Available online at https://s3.amazonaws.com/cdn.smfm.org/publications/224/download-491f0e6962960848d2097447ab57a024.pdf. Accessed 12/26/2016.

(2016 September). American College of Obstetricians and Gynecologists. Screening for birth defects.  Available online at http://www.acog.org/~/media/For%20Patients/faq165.pdf?dmc=1&ts=20130213T1410405774. Accessed 12/26/2016.

(2016 October). March of Dimes. Down syndrome. Available online at http://www.marchofdimes.com/baby/birthdefects_downsyndrome.html. Accessed 12/26/2016.

(Reviewed 2016 October 20). Centers for Disease Control and Prevention. Birth defects, diagnosis. Available online at http://www.cdc.gov/ncbddd/birthdefects/diagnosis.html. Accessed 12/26/2016.

(© 1995-2016). Mayo Medical Laboratories. Quad screen (second trimester) maternal, serum. Available online at http://www.mayomedicallaboratories.com/test-catalog/Clinical+and+Interpretive/81149. Accessed 12/26/2016.

(© 2016). Trisomy 18 Foundation. What is trisomy 18? Available online at http://www.trisomy18.org/site/PageServer?pagename=whatisT18_whatis. Accessed 12/26/2016.

(© 2016). Quest Diagnostics Test Center. Prenatal Screening and Diagnosis of Neural Tube Defects, Down Syndrome, and Trisomy 18. Available online at http://www.questdiagnostics.com/testcenter/testguide.action?dc=CF_PrenatScreen. Accessed January 2017.

(September 2016) American Congress of Obstetricians and Gynecologists. Prenatal Genetic Screening Tests. Available online at http://www.acog.org/Patients/FAQs/Prenatal-Genetic-Screening-Tests?IsMobileSet=false. Accessed January 2017.

Sources Used in Previous Reviews

Thomas, Clayton L., Editor (1997). Taber's Cyclopedic Medical Dictionary. F.A. Davis Company, Philadelphia, PA [18th Edition].

Pagana, Kathleen D. & Pagana, Timothy J. (2001). Mosby's Diagnostic and Laboratory Test Reference 5th Edition: Mosby, Inc., Saint Louis, MO.

Shirley L. Welch, PhD, DABCC, FACB. Director of Chemistry, Kaiser Permanente, NW, Portland, OR.

Barkai G, Goldman B, Ries L, Chake R, Zer T, Cuckle H. Prenatal Diagnosis 13: 843-850 (1993).

Lambert-Messerlian GM, Saller DN, Tumber MB, French CA, Peterson CJ, Canick JA. Prenatal Diagnosis 18: 1061-1067 (1998).

Cuckle H, et al. Prenatal Diagnosis 10: 71-77 (1990).

(2006 January). Healthy Pregnancy, The Second Trimester. WomensHealth.gov [on-line information]. Available online at http://www.womenshealth.gov/pregnancy/pregnancy/2nd.cfm. Accessed 2/25/07.

Barclay, L. and Lie, D. (2007 January 4). New Guidelines Recommend Universal Prenatal Screening for Down Syndrome. Medscape Medical News [on-line CME]. Available online at http://www.medscape.com/viewarticle/550256. Accessed 2/25/07.

Pagana, Kathleen D. & Pagana, Timothy J. (2007). Mosby's Diagnostic and Laboratory Test Reference 5th Edition: Mosby, Inc., Saint Louis, MO. Pp. 47-49, 416-419, 651-652.

Ashwood, E. et. al., Reviewed (2007 January). Prenatal Screening and Diagnosis. ARUP Consult [on-line information]. Available online at http://www.arupconsult.com/Topics/Perinatal_Disease/Prenatal_Screening_and_Diagnosis.html. Accessed 2/17/07.

Tietz Textbook of Clinical Chemistry and Molecular Diagnostics. Burtis CA, Ashwood ER, Bruns DE, eds. St. Louis: Elsevier Saunders; 2006, Pp 2167-2173.

(April 9, 2010) Singh D. Prenatal Diagnosis for Congenital Malformations and Genetic Disorders. Medscape article. Available online at http://emedicine.medscape.com/article/1200683-overview. Accessed September 2010.

(2008 November). Maternal Blood Screening for Birth Defects. March of Dimes [On-line information]. Available online at http://www.marchofdimes.com/professionals/14332_1166.asp. Accessed September 2010.

Clarke, W. and Dufour, D. R., Editors (2006). Contemporary Practice in Clinical Chemistry. AACC Press, Washington, DC, Pp 416-418.

(September 2009) American Pregnancy Association. Triple Screen Test: Multiple Marker Screen and Quad Screen. Available online at http://www.americanpregnancy.org/prenataltesting/. Accessed September 2010.

(June 30, 2010) Mayo Clinic. Quad Screen. Available online at http://www.mayoclinic.com/health/quad-screen/MY00127. Accessed September 2010.

Trisomy 18 Foundation. Available online at http://www.trisomy18.org. Accessed September 2010.

Barclay, L. and Lie, D. (Reviewed 2008 January 3). New Guidelines Recommend Universal Prenatal Screening for Down Syndrome. Medscape Medical News [on-line CME]. Available online at http://www.medscape.com/viewarticle/550256. Accessed September 2010.

Trisomy 18 Foundation. What is Trisomy 18? Available online at http://www.trisomy18.org/site/PageServer?pagename=whatisT18_whatis. Accessed February 2013.

March of Dimes. Birth defects: Down syndrome. Available online at http://www.marchofdimes.com/baby/birthdefects_downsyndrome.html. Accessed February 2013.

Centers for Disease Control and Prevention. Folic Acid Recommendations. Available online at http://www.cdc.gov/ncbddd/folicacid/recommendations.html. Accessed February 2013.

Centers for Disease Control and Prevention. Birth Defects, Diagnosis. Available online at http://www.cdc.gov/ncbddd/birthdefects/diagnosis.html. Accessed February 2013.

ACOG. Screening for Birth Defects. Available online at http://www.acog.org/~/media/For%20Patients/faq165.pdf?dmc=1&ts=20130213T1410405774. Accessed February 2013.

KidsHealth.org. What is the Multiple Marker Test? Available online at http://kidshealth.org/parent/system/medical/triple_screen.html. Accessed February 2013.

MayoClinic.com. Quad screen. Available online at http://www.mayoclinic.com/health/quad-screen/MY00127. Accessed February 2013.

ARUP Laboratories. Maternal Serum Screen, Alpha Fetoprotein, hCG, and Estriol. Available online at http://www.aruplab.com/guides/ug/tests/0080108.jsp. Accessed February 2013.

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