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The Sister Study

Baseline Specimen collection

Baseline Specimen Collection and Processing

A printable version of the Sister Study Baseline Specimen Collection and Processing Summary is available here:
Sister Study Baseline Specimen Collection and Processing Summary
(Requires Adobe Acrobat - Download Here) exit disclaimer

Sister Study specimens were collected at study participants' homes or a mutually agreed upon alternate site, such as a doctor’s office. Participants were sent a kit for collecting first morning void urine, toenails, and household dust. Samples, along with anthropometric and blood pressure measurements, were collected by examiners from a national in‐home phlebotomy service.

Baseline
Specimen N % of Cohort
Blood

50,433

99.1%

Saliva

354

0.7%

Urine

50,705

99.7%

Toenails

49,835

97.9%

Dust

50,367

99.0%

DNA has been extracted from baseline blood samples for a random subcohort (~4% of Sister Study cohort), breast and ovarian cancer cases as they accrue, and other studies as needed. To see the current number of participants with DNA extracted, click here: Sister Study Participants with DNA Extracted. (Requires Adobe Acrobat - Download Here) exit disclaimer

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Baseline Specimen Collection

Blood collection: Participants were instructed to fast for at least eight hours prior to their blood draw. The examiner drew the blood as shown in the table below, and retrieved the urine and environmental samples. Each participant had approximately 45 ml of blood collected into six BD Vacutainer® tubes, including two EDTA tubes, two serum tubes and two ACD-B tubes.

Tube Color/Top Tube Type Draw Order Packing Special Handling
Red

10 ml

(1)

Cold *

*Only while in transit back to exam office before processing. Invert

Red

10 ml

(2)

Cold *

*Only while in transit back to exam office before processing. Invert

Purple

EDTA
10 ml

(3)

Ambient

Invert

Yellow

ACD-B
6 ml

(4)

Ambient

Invert

Tan

EDTA
3 ml

(5)

Ambient

Invert

Yellow

ACD-B
6 ml

(6)

Ambient

Invert

 

Saliva collection: In the event that a blood sample could not be collected due to an unsuccessful phlebotomy, participants were asked to do a saliva collection using an Oragene™ DNA self-collection saliva kit. Although this has its limitations compared to blood, it provides a DNA source for genetic analyses.

Urine collection: On the morning of the examiner visit, participants collected approximately 60 ml of their first urine of the day in a collection cup that was pre-tested for phthalates.

Toenail collection: After removing any nail polish, the participant took a clipping from each toenail on both feet. Big toenails were stored separately from other toenails in provided envelopes.

Dust collection: Three door frames from three different rooms in the house were swiped for dust resulting in six dirty swabs from three different locations.

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Baseline Specimen Processing

Serum and blood clot: Two red-top tubes were centrifuged in the field immediately after collection in order to isolate the serum from the red cells as soon as possible. The serum was transferred to a standard transport tube for return to the lab. Upon arrival, the serum was stored in approximately fifteen 0.5 ml CryoBioSystem™ (CBS™) straws in liquid nitrogen vapor phase. One blood clot was stored in a cryovial.

Plasma and EDTA whole blood: A 10.0 ml EDTA BD Vacutainer® tube (purple top) provided whole blood, plasma, and dried blood. A 1.0 ml aliquot of whole blood was stored in a cryovial and two types of dry blood storage cards were spotted and stored (60 µl per spot). One was a treated card chemically impregnated to lyse cells and stabilize DNA and the other was an untreated card. The remaining whole blood was centrifuged and the plasma was isolated and stored in 0.5 ml CBS™ straws in liquid nitrogen vapor phase. A 3.0 ml EDTA BD Vacutainer® tube (tan top) was stored in the original Vacutainer® tube at -20°C for future analysis of metals, trace elements, and environmental contaminants.

ACD-B whole blood: One of the ACD-B Vacutainer® tubes was aliquotted into six 1.0 ml cryovials and cryopreserved with 10% DMSO using a programmable freezer. Fifteen percent of the time, this ACD-B tube was selected for lymphocyte isolation as an alternative protocol. Selection was based on an algorithm that predetermined a high risk group based on age of enrollment and the affected sister’s age at diagnosis plus randomly selected controls. The second ACD-B tube was aliquotted into one 4.5 ml cryovial and two 1.0 ml cryovials and stored in liquid nitrogen vapor phase.

Urine: Immediately upon receipt at the Sister Study lab, a basic chemistry urinalysis was performed to measure protein, creatinine, blood, leukocytes, nitrite, glucose, ketone, pH, and specific gravity. Urine was aliquotted into twenty 0.5 ml CBS™ straws, five 1.0 ml vials, and one 3.6 ml vial. Urine straws were stored in liquid nitrogen vapor phase and vials were stored in -80°C mechanical freezers.

Specimen Storage Summary per Sister Study Participant
Material type Additive Container Volume Final Storage Temp.

Blood clot

None

Vial

1

-80C/LN

Lymphocytes

ACD

Cryopreserved vial

1-2 vials, ~3 million cells/vial

LN

Plasma

EDTA

Straws

9 X 0.5 ml

LN

Serum

None

Straws

17 X 0.5 ml

LN

Whole blood

ACD

Vial

1 X 4.5 ml

-80C/LN

Whole blood

ACD

Vial

2 X 1.0 ml

LN

Whole blood

ACD

Cryopreserved vial

6 X 1.0 ml

LN

Whole blood

EDTA

Vacutainer

1 X 3.0 ml

-20C

Whole blood

EDTA

Vial

1 X 1.0 ml

-80C

Whole blood

EDTA

Filter cards

2

Ambient

Urine

None

Straws

20 X 0.5 ml

LN

Urine

None

Vial

1 X 3.6 ml

-80C

Urine

None

Vial

5 X 1.0 ml

-80C

Toenails

None

Envelope

2

Ambient

Household dust

None

Alcohol swabs in sealed plastic bag

6 swabs

-20C


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