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Epic Test Code LAB560 Quad Screen (Second Trimester) Maternal, Serum

Additional Codes

MML:QUAD1


Specimen Required


Container/Tube: 

Preferred: Red top

Acceptable: Serum gel

Specimen Volume: 1 mL

Collection Instructions:

1. Do not draw specimen after amniocentesis, as this could affect results.

2. Centrifuge Immediately

Additional Information:

1. For an assessment that includes neural tube defect results, gestational age must be between 15 weeks, 0 days and 22 weeks, 6 days.

2. Assessments for trisomy 21 (Down syndrome) and trisomy 18 (Edwards syndrome) only are available between 14 weeks, 0 days and 22 weeks, 6 days.

3. Initial or repeat testing is determined in the laboratory at the time of report and will be reported accordingly. To be considered a repeat test for the patient, the testing must be within the same pregnancy and trimester, with interpretable results for the same tests, and both tests are performed at Mayo Clinic.

4. Patient education brochure (T522) is available upon request.


Forms

1. Second Trimester Maternal Screening Alpha-Fetoprotein (AFP)/QUAD Screen Patient Information (T595) is required in Special Instructions

2. If not ordering electronically, complete, print, and send a General Request (T239) with the specimen.

Useful For

Prenatal screening for open neural tube defect (alpha-fetoprotein only), trisomy 21 (alpha-fetoprotein, human chorionic gonadotropin, estriol, and inhibin A) and trisomy 18 (alpha-fetoprotein, human chorionic gonadotropin, and estriol)

Testing Algorithm

See Prenatal Aneuploidy Screening and Diagnostic Testing Options in Special Instructions

Method Name

Immunoenzymatic Assay

Reporting Name

QUAD SCRN (2nd Tri) Maternal, S

Specimen Type

Serum

Specimen Minimum Volume

0.75 mL

Specimen Stability Information

Specimen Type Temperature Time
Serum Refrigerated (preferred) 7 days
  Frozen  90 days

Reject Due To

Hemolysis

Mild OK; Gross reject

Lipemia

Mild OK; Gross OK

Icterus

NA

Other

NA

Clinical Information

Multiple marker serum screening has become a standard tool used in obstetrical care to identify pregnancies that may have an increased risk for certain birth defects, including neural tube defects (NTD), trisomy 21 (Down syndrome), and trisomy 18 (Edwards syndrome). The screen is performed by measuring analytes in maternal serum that are produced by the fetus and the placenta. The analyte values along with maternal demographic information such as age, weight, gestational age, diabetic status, and race are used together in a mathematical model to derive a risk estimate. The laboratory establishes a specific cutoff for each condition, which classifies each screen as either screen-positive or screen-negative. A screen-positive result indicates that the value obtained exceeds the established cutoff. A positive screen does not provide a diagnosis, but indicates that further evaluation should be considered.

 

Analytes:

Alpha-Fetoprotein (AFP)

AFP is a fetal protein that is initially produced in the fetal yolk sac and liver. A small amount also is produced by the gastrointestinal tract. By the end of the first trimester, nearly all of the AFP is produced by the fetal liver. The concentration of AFP peaks in fetal serum between 10 to 13 weeks. Fetal AFP diffuses across the placental barrier into the maternal circulation. A small amount also is transported from the amniotic cavity.

 

The AFP concentration in maternal serum rises throughout pregnancy, from a nonpregnancy level of 0.2, to about 250 ng/mL at 32 weeks gestation. If the fetus has an open NTD, AFP is thought to leak directly into the amniotic fluid causing unexpectedly high concentrations of AFP. Subsequently, the AFP reaches the maternal circulation, thus producing elevated serum levels. Other fetal abnormalities such as omphalocele, gastroschisis, congenital renal disease, esophageal atresia, and other fetal distress situations such as threatened abortion, and fetal demise also may show AFP elevations. Increased maternal serum AFP values also may be seen in multiple pregnancies and in unaffected singleton pregnancies in which the gestational age has been underestimated.

 

Lower maternal serum AFP values have been associated with an increased risk for genetic conditions such as trisomy 21 (Down syndrome) and trisomy 18.

 

Estriol (uE3)

Estriol, the principal circulatory estrogen hormone in the blood during pregnancy, is synthesized by the intact feto-placental unit. Estriol exists in maternal blood as a mixture of the unconjugated form and a number of conjugates. The half-life of unconjugated estriol in the maternal blood system is 20 to 30 minutes because the maternal liver quickly conjugates estriol to make it more water soluble for urinary excretion. Estriol levels increase during the course of pregnancy. Decreased unconjugated estriol has been shown to be a marker for trisomy 21 and trisomy 18. Low levels of estriol also have been associated with overestimation of gestation, pregnancy loss, Smith-Lemli-Opitz, and X-linked ichthyosis (placental sulfatase deficiency).

 

Human Chorionic Gonadotropin (Total Beta-hCG: ThCG)

hCG is a glycoprotein consisting of 2 noncovalently bound subunits. The alpha subunit is identical to that of luteinizing hormone (LH), follicle-stimulating hormone (FSH), and thyroid-stimulating hormone (TSH), while the beta subunit has significant homology to the beta subunit of LH and limited similarity to the FSH and TSH beta subunits. The beta subunit determines the unique physiological, biochemical, and immunological properties of hCG.

 

The CGA gene (glycoprotein hormones, alpha polypeptide) is thought to have developed through gene duplication from the LH gene in a limited number of mammalian species. hCG only plays an important physiological role in primates (including humans), where it is synthesized by placental cells, starting very early in pregnancy, and serves to maintain the corpus luteum, and hence, progesterone production, during the first trimester. Thereafter, the concentration of hCG begins to fall as the placenta begins to produce steroid hormones and the role of the corpus luteum in maintaining pregnancy diminishes.

 

Increased total hCG levels are associated with trisomy 21, while decreased levels may be seen in trisomy 18. Elevations of hCG also can be seen in multiple pregnancies, unaffected singleton pregnancies in which the gestational age has been overestimated, triploidy, fetal loss, and hydrops fetalis.

 

Inhibin A

Inhibins are a family of heterodimeric glycoproteins, primarily secreted by ovarian granulosa cells and testicular Sertoli cells, which consist of disulfide-linked alpha and beta subunits. While the alpha subunits are identical in all inhibins, the beta subunits exist in 2 major forms, termed A and B, each of which can occur in different isoforms. Depending on whether an inhibin heterodimer contains a beta A or a beta B chain, they are designated as inhibin A or inhibin B, respectively. Together with the related activins, which are homodimers or heterodimers of beta A and B chains, the inhibins are involved in gonadal-pituitary feedback and in paracrine regulation of germ cell growth and maturation. During pregnancy, inhibins and activins are produced by the feto-placental unit in increasing quantities, mirroring fetal growth. Their physiological role during pregnancy is uncertain. They are secreted into the coelomic and amniotic fluid, but only inhibin A is found in appreciable quantities in the maternal circulation during the first and second trimesters.

 

Maternal inhibin A levels are correlated with maternal hCG levels and are abnormal in the same conditions that are associated with abnormal hCG levels (eg, inhibin A levels are typically higher in trisomy 21 pregnancies). However, despite their similar behavior, measuring maternal serum inhibin A concentrations in addition to maternal serum hCG concentrations further improves the sensitivity and specificity of maternal multiple marker screening for trisomy 21.

Reference Values

NEURAL TUBE DEFECTS

An AFP multiple of the median (MoM) <2.5 is reported as screen negative. AFP MoMs ≥2.5 (singleton and twin pregnancies) are reported as screen positive. 

 

DOWN SYNDROME

Calculated screen risks <1/270 are reported as screen negative, risks ≥1/270 are reported as screen positive.

 

TRISOMY 18

Calculated screen risks <1/100 are reported as screen negative, risks ≥1/100 are reported as screen positive.

 

An interpretive report will be provided.

Interpretation

Neural Tube Defects (NTD):

A screen-negative result indicates that the calculated alpha-fetoprotein (AFP) multiple of the median (MoM) falls below the established cutoff of 2.50 MoM. A negative screen does not guarantee the absence of NTD.

 

A screen-positive result indicates that the calculated AFP MoM is 2.50 MoM or greater, and may indicate an increased risk for open NTD. The actual risk depends on the level of AFP and the individual's pretest risk of having a child with NTD based on family history, geographical location, maternal conditions such as diabetes and epilepsy, and use of folate prior to conception. A screen-positive result does not infer a definitive diagnosis of NTD, but indicates that further evaluation should be considered. Approximately 80% of pregnancies affected with NTD have elevated AFP, MoM values greater than 2.5.

 

Trisomy 21 (Down syndrome) and Trisomy 18 (Edwards syndrome):

A screen-negative result indicates that the calculated screen risk is below the established cutoff of 1/270 for trisomy 21 and 1/100 for trisomy 18. A negative screen does not guarantee the absence of trisomy 21 or trisomy 18.

 

When a trisomy 21 second-trimester risk cutoff of 1/270 is used for follow-up, the combination of maternal age, AFP, estriol, hCG, and inhibin A has an overall detection rate of approximately 77% to 81% with a false-positive rate of 6% to 7%. In practice, both the detection rate and false-positive rate increase with age. The detection rate ranges from 66% (early teens) to 99% (late 40s), with false-positive rates of between 3% and 62%, respectively.

 

The detection rate for trisomy 18 is 60% to 80% using a second trimester cutoff of 1/100.

 

Follow-up:

Upon receiving maternal serum screening results, all information used in the risk calculation should be reviewed for accuracy (maternal date of birth, gestational dating, etc.). If any information is incorrect, the laboratory should be contacted for a recalculation of the estimated risks.

 

Screen-negative results typically do not warrant further evaluation.

 

Ultrasound is recommended to confirm dates for NTD or trisomy 21 screen-positive results. Many pregnancies affected with trisomy 18 are small for gestational age. Recalculations that lower the gestational age may decrease the detection rate for trisomy 18. If ultrasound yields new dates that differ by at least 7 days, a recalculation should be considered. If dates are confirmed, high-resolution ultrasound and amniocentesis (including amniotic fluid AFP and acetylcholinesterase measurements for NTD) are typically offered.

Cautions

Variables Affecting Marker Levels:

Race, weight, smoking, multiple fetus pregnancy, insulin-dependent diabetes (IDD), and in vitro fertilization (IVF) may affect marker concentrations. Black mothers tend to have higher alpha-fetoprotein (AFP) levels but lower risk of neural tube defects (NTD) and are assigned to a separate AFP median set. All multiple of the median (MoMs) are adjusted for maternal weight (to account for dilution effects in heavier mothers). The AFP, estriol (uE3), and inhibin MoMs are adjusted upward in IDD to account for lower values in diabetic pregnancies. Human chorionic gonadotropin (hCG) levels are higher and uE3 levels are lower in pregnancies conceived by IVF, MoMs are adjusted accordingly to account for the alterations. Smoking results in higher second trimester maternal serum AFP and inhibin A levels and lower uE3 and hCG levels. MoMs are adjusted accordingly to account for analyte differences in smokers.

 

The estimated risk calculations and screen results are dependent on accurate information for gestation, maternal age, race, IDD, and weight. Inaccurate information can lead to significant alterations in the estimated risk. In particular, erroneous assessment of gestational age can result in false-positive or false-negative screen results. Because of its increased accuracy, we therefore recommend determination of gestational age by ultrasound, rather than by last menstrual period, when possible.

 

A screen-negative result does not guarantee the absence of fetal defects. A screen-positive result does not provide a diagnosis, but indicates that further diagnostic testing should be considered (an unaffected fetus may have screen-positive result for unknown reasons).

 

Valid measurements of AFP in maternal serum cannot be made after amniocentesis.

 

Triplet and higher multiple pregnancies cannot be interpreted.

 

Each center offering maternal serum screening to patients should establish a standard screening protocol, which provides pre- and post-screening education and appropriate follow-up for screen-positive results.

Day(s) and Time(s) Performed

Monday through Friday; 8 a.m.-4:30 p.m.

Analytic Time

1 day

Specimen Retention Time

12 months

Performing Laboratory

Mayo Clinic Laboratories in Rochester

CPT Code Information

81511

NY State Approved

Yes