Alzheimer’s Disease in Down Syndrome (ADDS) in collaboration with The National Centralized Repository for Alzheimer’s Disease and Related Dementias (NCRAD) Blood-Based Biospecimen Training Slides
Contact I Information • Questions? Please contact NCRAD Coordinators at: • Phone: 1-800-526-2839 • E-mail: alzstudy@iu.edu • Website: www.ncrad.org
Traini ning O g Over ervi view: • Blood-Based Collection Schedule • Kit Request Module • Specimen Labels • Handling/Processing Study Specimens • Sample Shipping • NCRAD Website • Questions?
ADDS DDS Bl Blood Ba Based Co Collection Schedu edule: e: Baseline 3-Month 16-Month 19-Month 32-Month 35-Month (1A) (1B) (2A) (2B) (3A) (3B) Serum X X X DNA X X X Plasma X X X
Ki Kit Req eques est M Mod odule http://kits.iu.edu/adds-niad
Kit R Req eques est M Modu odule • An initial stock of kits will be delivered prior to the designated site specific start date. • Kits and individual supplies are available to order: • ADDS Blood Kit • ADDS Frozen Blood Shipping Supply Kit • Blood Supplemental Kit • CSF Supplemental Supply Kit • Lumbar Puncture Trays • Frozen CSF Shipping Supply Kit
NCRAD AD K Kit R Request Module 1. Choose your site from the drop down list 2. The coordinator name and contact information will appear 3. Verify that this information is accurate, correct if necessary
ADDS a S and nd NiAD Kits A Available e
St Study V Visi sit K Kits • Indicate the quantity needed of each kit 1 • Once selected, kit components of the chosen kit will appear at the bottom of the screen (Pictured) • Click “Submit” to turn in your request. • The IU staff will notify you that your request has been received and address any issues. • **Note: You can order more than one type of kit in a single kit request**
Sp Speci ecimen L Lab abels
Label bel T Type Su e Summary 1. Kit Number Labels 2. Site and Subject ID Labels 3. Collection and Aliquot Tube Labels • Differ by specimen type
Kit Number er L Label bels • Used to track patient samples and provide quality assurance • Will be placed on the following locations: 1. Biological Sample and Shipment Notification Form 2. Outside cryobox that houses aliquot tubes during storage and shipment Provided by NCRAD in the kits
Si Site a e and nd Su Subjec ect I ID Label bel • Subjects will be identified by their site ID and subject ID • The subject ID may only be available shortly before the visit • Sites will be responsible for handwriting this onto the provided labels • Must use fine point permanent marker • Each site will receive 4 markers in initial kit supply
Si Site a e and nd Su Subjec ect I ID Label bel Con Cont. • Write information on label prior to adhering to tube • Label will be placed on all collection tubes • (2) Serum Separator (Gold-Top) Blood Collection Tube (5 ml) • EDTA (Lavender-Top) Blood Collection Tube (10 ml) • Kits will include one extra label
Col Collec ection a and Aliquot T Tube L e Label bels Specimen Number (assigned by NCRAD) Repository Name Sample Type Kit # (assigned by NCRAD) unique to the subject and visit
Collec ection a on and d Aliquot quot T Tube L e Label els- Bl Blood • Labels to be placed on ALL collection and aliquot tubes 1. Serum Separator (Gold-Top) Blood Collection Tube (x2) • Serum aliquots 2. EDTA (Lavender-Top) Blood Collection Tube (x1) • Plasma aliquots • Buffy coat aliquot
Col ollec ection Tube ubes – Blood od Collection/Aliquot tube label *place barcode near top Site and Subject ID Label EDTA (Lavender-Top) Serum Separator Blood Collection Tube (Gold-Top) Blood (10 ml) Collection Tube (5 ml)
Col ollec ection Tube ubes - Blood od Label 2: Site and ID Label Label 1: Collection Tube Label EDTA Tube Serum Separator Tubes • All collection tubes will have two labels • The Collection Tube Labels • The handwritten Site and Subject ID Label
Aliquot T Tube L e Label bels – Ser Serum, P Plasma and nd Buf Buffy Coa Coat • Collection and Aliquot tube label only • Please place barcode near cap
NCRA CRAD Ki Kit R Req eques est Mod odule: e: Whe hen It M Mus ust be U be Used ed • Each site will be responsible for ordering kits (labels included) and maintaining a supplies on site for scheduled participants • To order, sites will use the Indiana University online kit ordering module: http://kits.iu.edu/adds-niad • Allow a minimum of 2 weeks for your order to be processed and delivered.
Han andling/Proce ocessi ssing g St Study Sp Speci ecimens s
Site R Si e Requ equired E Equ quipment Blood Collection/Safety Processing/Storage Equipment Equipment 1. PPE 1. Centrifuge capable of ≥2000 rcf with refrigeration to 4⁰C • Lab Coat, Safety Glasses 2. - 80⁰C Freezer 2. Tourniquet 3. Wet Ice Bucket 3. Alcohol Prep Pad 4. Gauze Pad 5. Butterfly Needles 6. Bandage 7. Sharps Bin and Lid
Draw O Order ***Important Note*** In order to ensure the highest quality samples are collected, processed, and stored, it is essential to follow the specific collection, processing, and shipment procedures detailed in the following pages. Please read the following instructions first before collecting any specimens. Have all your supplies and equipment out and prepared prior to drawing blood. Draw blood in the following order: 1. Serum Separator (Gold-Top) Blood Collection Tube (5 ml) for Serum x 2 2. EDTA (Lavender-Top) Blood Collection Tube (10 ml) for DNA and Plasma
Sa Sample Col e Collec ection - Blood Number Tube Type of Tubes Tube Image Drawn 1. Serum Separator (Gold-Top) x2 Blood Collection Tube (5 ml) 2. EDTA (Lavender-Top) x1 Blood Collection Tube (10 ml)
Aliqu quot C Cap C Colors
Serum Preparation (5 ml Gold-Top Tube) Step Six Serum Serum Serum Serum Serum Serum Serum Serum Serum • Adhere preprinted labels to the clear cap • • Using a clean transfer pipette, Collect blood cryovials with red stickers Immediately transfer Serum from both 5 mL Gold- • in (2) 5 mL Aliquot 0.25 ml into each cryovial tube. after blood Top tubes to the 15 mL conical tube. • Gold-Top If a residual aliquot is created, document draw, invert • Mix the 15 mL conical tube gently by tubes specimen number and volume on Sample tube 5 times inverting 3-4 times. allowing Notification Form. to mix blood to flow • samples. Store serum aliquots at -80°C until for 10 shipment. seconds and ensure blood flow has stopped.
Serum um S Sepa parator or T Tube ( e (Serum um C Collec ection) on) SERUM GEL MATRIX BLOOD CLOT Serum Separator Tube Serum Separator Tube after Immediately after Blood Draw Centrifuge Create 16 to 21 aliquots of 0.25ml; if residual aliquot created, document specimen number and volume on sample form
Plasma and Buffy Coat Preparation (10ml Lavender-Top Tube) Step Step Step Step Step Step Six One Three Four Two Five • Adhere preprinted labels to the clear cap cryovials with lavender stickers. • Aliquot 0.25 ml into each cryovial tube. • Store plasma aliquots at -80°C until shipment. Step • • • Place • • Collect blood Immediately Preferably within Store tubes at Immediately Seven thoroughly in Plasma after blood 30 minutes of room after blood • Adhere preprinted labels to the mixed tube on Tube allowing draw, invert draw, invert blood draw, temperature. blue cap cryovial. blood to flow tubes 8-10 wet ice until centrifuge samples • tube 5 times Label tubes with • Using a clean pipette tip, collect for 10 times to mix centrifugation at 2000 x g for 10 to mix preprinted labels the buffy coat (may have residual begins. seconds and samples. samples. minutes. prior to blood plasma and some RBCs included). • ensuring Samples need to be draw. • Transfer the buffy coat into the blood flow spun, aliquoted, cryovial tube. has stopped. and in the freezer • Store buffy coat aliquot at -80°C within 2 hours until shipment. from the time of collection.
EDTA TA Tube ( e (Plasma Col Collec ection) Create 16 to 21 aliquots of 0.25ml; if residual aliquot created, document specimen number and volume on sample form
EDTA TA Tube (Buffy C y Coat Collection on) Important Note: Buffy Coat aliquots will be distinguished from the plasma aliquots through a blue cap.
Sam Sample Sh e Shipping
Blood S Sample S Shipment S t Summary ADDS Processing/ Aliquoting Tubes to NCRAD Ship Sample Type Whole blood (Gold- 0.25 ml serum aliquots per 0.5 ml siliconized Top SST) for isolation Yes 16-21 Frozen of serum cryovial (clear caps with RED stickers) 0.25 ml plasma aliquots per 0.5 ml siliconized Whole blood Yes 16-21 Frozen cryovial (clear caps (lavender-Top EDTA) with lavender stickers) for isolation of plasma & buffy coat (for DNA extraction) 1 ml buffy coat aliquot Yes per 2.0 ml cryovial 1 Frozen (BLUE cap)
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