VAlidation of a Lower Cost aneUploidy scrEen

Women & Infants Hospital of Rhode Island (WIH) will serve as the Study Center and the
Laboratory Site. Enrollment Sites will secure local IRB approval, identify, consent and
enroll pregnant women, ship plasma samples to WIH and collect outcome information. Eligible
patients will be from one of two groups:

- The 'high risk' (HR) group - 250 women considering diagnostic testing, nearly all
following a positive cell free (cf)DNA screening test. At least one case of Down
syndrome should be detected for every three women enrolled, in addition to an occasional
case of T18 or T13.

- The 'low risk' (LR) group - 2,400 women having conventional cfDNA screening as their
primary screening test (i.e., no serum/ultrasound testing), representing the general
pregnancy population. Their group risk of Down syndrome should be about 1:500. This
group should have no more than 20% women age 35 and older. (Women whose conventional
cfDNA screen is positive would then be eligible for a second enrollment in the 'high
risk' group).

Both groups of women will be asked to provide three full 10mL Streck tubes of blood (two
tubes minimum), agree to release limited clinical information, provide signed consent
(including for FDA personnel to review records) and consent to have Enrollment Site staff
review and report needed information from newborn/infant examination records, if requested.
Identifiable patient information will be held at Enrollment Sites and not released to the
Laboratory or Study Center. Sample and outcome information will be identified only by a
unique study code. Individual cfDNA test results will not be returned to the Enrollment
Sites, providers, or enrolled women. Enrollment Sites will receive a summary of results once
the study ends.

Enrollment Sites are likely to enroll either HR or LR women, although a few sites may be able
to enroll both. Both settings will have genetic counselors, physicians, research assistants
or other medical staff available to identify, inform, consent and enroll women, as well as to
collect and ship samples and obtain pregnancy outcome information. Staff designated as
participating in the VALUE Study will be qualified to inform eligible women of the study's
benefits and harms, and collect/document informed consent (which will remain at that site).
The Study Center may request confirmation of consent, should it become necessary. Outcomes
for HR women choosing diagnostic testing will be the results of the karyotype; those
declining diagnostic testing will have newborn/infant information collected( often a newborn
karyotype). In the LR population, a negative cfDNA test result will be accepted as ruling out
a common autosomal trisomy, as the residual risk would be very low (<1:20,000 to < 1:50,000).
For LR women with a positive cfDNA test, results of diagnostic testing would be requested. If
diagnostic test results are unavailable, a review of the newborn examination/karyotype will
be required for confirmation. Any conflicting results between the clinical tests and the
VALUE Study results(false positive / false negative) would also require resolution that may
involve review of newborn/infant records.

The VALUE Study intends to enroll approximately 2,400 LR women through as many as 8 LR
Enrollment Sites associated with general risk obstetrical care practices. The 2,400 target is
set to: 1) provide a reasonably confident estimate of an expected low false positive rate
(e.g., 0.2%) and 2) provide sufficient numbers to exercise the testing platform over a 12
month time period. The target of 2,400 is large enough to meet both objectives and is
slightly more than the largest number of euploid samples tested in any of the original NGS HR
cfDNA validation studies (2011-2012). The number of trisomies detected in this LR group
should be fewer than 10.An estimated 2% (48) of these LR women will have a failed/no call
cfDNA test..

Sample handling:

Plasma samples could be tested fresh or stored and tested once the assay system has been
validated. Outcome information will not be collected from Enrollment Sites until samples have
been tested, ensuring blinding of laboratory to outcomes.

Timeframe:

Identification of Enrollment Sites and securing Investigational Review Board (IRB) approval
will take three months. Efforts related to set-up and qualification of the testing laboratory
is expected to take four to six months as will study-specific software development. Active
enrollment is scheduled for 12 months. The intent is to run the majority of samples fresh,
allowing for long-term assay variability to be studied. Follow-up will be completed, for the
most part, within two months of the final testing (except for the small proportion of women
who tested positive but did not have diagnostic testing). These will have outcomes confirmed
at the time of delivery.

Inclusion Criteria:

- At least 18 years old

- 10-20 weeks gestation

- Low Risk group

- no known aneuploidy risk

- scheduled for cfDNA screen as primary aneuploidy screen

- High Risk group

- positive cfDNA screen

- scheduled to discuss CVS/amniocentesis as confirmatory test

Exclusion Criteria:

- Triplet or higher order pregnancy

- Low Risk group

- positive nuchal translucency (NT) or abnormal ultrasound

- previous pregnancy with aneuploidy