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Reporting Title: Sequential Screen | FB Test Definition: SECTRISEQ Testing Location: Melville, NY Reporting Location: Melville, NY Description: Sequential Screen | FB includes measuring second trimester maternal serum alpha-fetoprotein (AFP), free Beta human chorionic gonadotropin (fbhCG), and inhibin A. These measurements are compared to median values for a given gestational age and a MoM is calculated for each. These results are combined with the first trimester results for PAPP-A, nuchal translucency and nasal bone and are entered into a multivariate algorithm that includes the mother's age to derive risk factors for Down syndrome, trisomy 18, and neural tube defects. The screen for NTD uses the AFP MoM only. Analytical Method(s): All assays are performed on a PerkinElmer AutoDELFIA instrument.

1. AFP is measured using PerkinElmer’s solid phase 2-site sandwich fluorometric assay (product number: PKI B079-201).

2. Free Beta hCG is measured using a lab-developed solid phase 2-site sandwich fluorometric assay.

3. UE3 is measured using PerkinElmer’s solid phase competition fluorometric assay. (product number: PKI B083-CO1)

4. Inhibin A is measured using PerkinElmer’s solid phase 2-site sandwich fluorometric assay (product number: PKI B064-112).

Patient preparations: Counsel patient on prenatal screening for aneuploidy and open neural tube defects. Specimen Requirements: Sequential maternal screening is a 2-part test that includes a first-trimester sample (First Trimester Screen| FB) and a second-trimester sample (Sequential Screen | FB). Do not order this test unless the First Trimester Screen| FB test has already been performed for the patient. If a standalone second-trimester screen is desired, order Quad Screen | FB. Container/Tube: Red-top Vacutainer® tube or Serum Separator Tube (Red/Grey or Gold Top SST) Specimen Volume: 0.5 ml of spun serum or 5 ml of unspun whole blood Specimen Stability: Serum samples are stable at ambient temperature for 6 days. Specimen Rejection Criteria: hemolysis, lipemia, incorrect tube type Specimen Collection Instructions See Blood Specimen Collection from Venipuncture Instruction Manual. Additional Information:

1. Indications for Testing: General population screening of pregnant women 2. Special Timing: Draw blood between 15 weeks, 0 days and 21 weeks, 6 days.

Document name: GEN-2-043-Attachment L-SequentialScreen | Fß

Instruction Manual

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3. Do not draw blood after performing amniocentesis, as that may lead to an artificially increased serum alpha-fetoprotein level and unreliable results.

4. Specimen Stability: Serum specimens are stable at ambient temperature for 6 days. CPT Code 1 x 82105, 1 x 84704, 1 x 82677, 1 x 86336 Reference Values: Down syndrome: After Screening Risk < 1 in300 are Within Range After Screening Risk > 1 in 300 are Increased Risk Trisomy 18 After Screening Risk < 1 in 150 are Within Range After Screening Risk > 1 in 150 are Increased Risk Total ONTDs/VWD AFP MoM < 2 MoM are Within Range AFP MoM 2-2.49 MoM are Borderline Elevation AFP MoM > 2.5 MoM are Significant Elevation An Interpretive Report will be provided. Supplemental Report: No Testing Algorithm: Follow up testing:

1. Increased Risk Results for Down syndrome or Trisomy 18 and GA<19 weeks: Genetic counseling and offer amniocentesis for diagnostic confirmation or noninvasive prenatal testing (NIPT).

2. Increased Risk Results for Down syndrome or Trisomy 18 and GA>=19 weeks: Genetic Counseling and offer of amniocentesis.

3. MSAFP Borderline Elevation and GA<19 weeks: Repeat blood or serum. 4. MSAFP Borderline Elevation and GA>=19 weeks: Genetic Counseling, Ultrasound and offer of

amniocentesis. 5. MSAFP Significant Elevation: Genetic Counseling, Ultrasound and offer of amniocentesis.

Consents/Authorizations: Patient signature on patient authorization/assignment on requisition form is required. Disclaimer: The test was developed and its performance characteristics determined by Eurofins NTD, LLC. It has not been cleared or approved by the U.S. Food and Drug Administration. The methods and performance characteristics have been reviewed and approved by the New York State Department of Health.

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Instruction Manual

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Test Requisition Instructions

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NTD Prenatal Screening Requisition Form

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EMR Ask at Order Entry (AOE) Questions: Test ID Question ID Description Type Required SECTRISEQ MATWT Maternal Weight Plain Text YES SECTRISEQ MWLBSKGS Units

LBS KGS

Answer List YES

SECTRISEQ ETHNIC Ethnicity African American/Caribbean Asian Asian Indian Caucasian Hispanic Native American Other

Answer List YES

SECTRISEQ HXCHROM Previous History of Chromosome Abnormality None Trisomy 21 Trisomy 18 Trisomy 13 Trisomy 21 and 18 Trisomy 21 and 13 Trisomy 21, 18, 13

Answer List YES

SECTRISEQ HXNTD Family History of ONTD None Prev Child Patient’s Sister’s Child Patient’s Brother’s Child Patient’s Sister Patient’s Brother Patient’s Aunt Patient’s Uncle Partner’s Prev Child Partner’s Brother

Answer List YES

SECTRISEQ IVFAGE IVF-Age of Egg (years) Plain Text NO SECTRISEQ NOF Number of Fetuses

1 2

Answer List YES

SECTRISEQ SMOKE Is Patient a Smoker No Yes

Answer List YES

SECTRISEQ VALPRO Valproic Acid or carbamazepine used during preg No

Answer List YES

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Yes SECTRISEQ IDDM Insulin dependent diabetic Prior to pregnancy

No Yes

Answer List YES

SECTRISEQ CO3MO Address-Country(If different 1st 3 mos. of preg) Plain Text NO SECTRISEQ ST3MO Address-State(If different 1st 3 mos. of preg) Plain Text NO EMR Result Codes: Data Type

Code LOINC Name Contains Result

Comments

CE RSKTBL Risk Table No Included if disorders are available ST DOWNS 59462-2 Down Syndrome Yes Risk result information contained in

NTEs ST T18 59462-2 Trisomy 18 Yes Risk result information contained in

NTEs ST TONTD 59462-2 Total ONTDs/VWD Yes Risk result information contained in

NTEs ST DOWNS-B 59462-2 Down Syndrome Twin B Yes Displayed for twin B, Risk result

information contained in NTEs ST T18 59462-2 Trisomy 18 Twin B Yes Displayed for twin B, Risk result

information contained in NTEs ST TONTD-B 59462-2 Total ONTDs/VWD Twin B Yes Risk result information contained in

NTEs CE MKRANA Markers/Analytes No Included if any 1T markers are

available CE 2T 2nd Trimester No CE AFP2 19176-7 AFP Yes Measurements contained in NTEs CE FBHCGV2 25373-2 Free Beta hCG Yes Measurements contained in NTEs CE UE3 15064-9 uE3 Yes Measurements contained in NTEs CE INH 23883-2 Inhibin-A Yes Measurements contained in NTEs CE 1T 1st Trimester No CE FBHCGV 25373-2 Free Beta hCG Yes Measurements contained in NTEs CE PAPPA 32046-5 PAPP-A Yes Measurements contained in NTEs CE AFP 19176-7 AFP Yes Measurements contained in NTEs CE PlGF PlGF Yes Measurements contained in NTEs CE INHIBIN 23883-2 Inhibin A Yes Measurements contained in NTEs CE NTV NT Yes Measurements contained in NTEs CE NTVB NT – Twin B Yes Displayed for twin B, Measurements

contained in NTEs ST NB Nasal Bone Yes ST NBB Nasal Bone – Twin B Yes Displayed for twin B CE DGD Demographic Data Yes Included if demographic data is

available (contained in NTEs) CE 2TD 2nd Trimester Data Yes Included if test-specific data is

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available (contained in NTEs) CE 1TD 1st Trimester Data Yes Included if test-specific data is

available (contained in NTEs) CE REC Recommendations Yes Only displayed if recommendations

are available CE REC-B Recommendations Twin B Yes Displayed for twin B, Only displayed if

recommendations are available CE COM 55107-7 Comments Yes Only displayed if comments are

available CE COM-B 55107-7 Comments Twin B Yes Displayed for twin B, Only displayed if

comments are available CE FTR Footer Yes Only displayed if footer is available CE FTR-B Footer Twin B Yes Displayed for twin B, Only displayed if

footer is available CE NOT Notification Yes Included for Unsatisfactory

Specimens Only CE NOT-B Notification Twin B Yes Included for Unsatisfactory

Specimens Only, Displayed for twin B

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Example Report

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Physician Information Brochure

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Patient Information Brochure

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