DBS英文版流程示意.pdf
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1、ACTN Laboratory Technologist Committee Version 1.1 ACTN Dried Blood Spots Procedure 11 March 2009 1 DRIED BLOOD SPOTS CARD COLLECTION, PROCESSING, AND STORAGE PROCEDURES 1. Rational 1.1 Dried blood spots (DBS) have been used for years for HIV-1 RNA, DNA, Genotyping, and other diagnostic assays. Drie
2、d blood spots can be used as an easy and inexpensive means for the collection and storage of peripheral blood specimens from infants, children and adults in settings where collection and storage of plasma is not optimal. Dried blood spots use a small amount of peripheral blood on protein saver filte
3、r paper cards from infant heel pricks, or finger sticks from adults and require less skill and supplies than venous blood draws. The cost and difficulties of cold- chain shipping of plasma samples is greatly reduced by the use of dried blood spots and they can be shipped as non-dangerous goods. 2. R
4、EAGENTS AND MATERIALS 2.1 Materials for Heel Stick Method 2.1.1 Unistick 2 device (Fisher Scientific #22-0227) 2.1.2 Alcohol swab 2.1.3 Dry sterile gauze pad 2.1.4 Whatman Protein Saver Card #903 (Whatman 10534612 Fisher Scientific # NC9307519) 2.1.5 Gas impermeable storage bag (Fisher Scientific# N
5、C9307519) 2.1.6 Desiccant pack (Whatman 10548234-Fisher Scientific# 50853571) 2.1.7 Humidity indicator Cards (Multisorb Des Manufacture # MS200032 Fisher Scientific # NC9511648) 2.1.8 Whatman card drying rack, (VWR catalogue # 89015-592) 2.1.9 Gloves preferably powder free 2.1.10 Water proof marker
6、2.2 Materials for Venous Blood Method Using Vacutainer Tubes 2.2.1 Vacutainer Evacuated Blood Collection Tubes 2.2.2 Alcohol swab 2.2.3 Tourniquet 2.2.3 Bandage/Plaster 2.2.4 Vacutainer Needle 2.2.5 Vacutainer Needle Holder 2.2.6 Whatman Protein Saver Card #903 (Whatman 10534612 Fisher Scientific #
7、NC9307519) 2.2.7 Gas impermeable storage bag (Fisher Scientific# NC9307519) 2.1.8 Desiccant pack (Whatman 10548234-Fisher Scientific# 50853571) ACTN Laboratory Technologist Committee Version 1.1 ACTN Dried Blood Spots Procedure 11 March 2009 2 2.1.9 Humidity indicator Cards (Multisorb Des Manufactur
8、e # MS200032 Fisher Scientific # NC9511648) 2.1.10 Whatman card drying rack (VWR catalogue # 89015-592) 2.1.11 Gloves preferably powder free 2.1.12 Water proof marker 3 Biosafety 3.1 It is essential that universal precautions be taken while working with these specimens. Dried blood spots on filter p
9、aper are not considered to be a biohazard. However, you should wear gloves and a lab coat/gown at all times, even with DBS, to ensure safe handling of samples. If you should tear a glove, remove the torn one and replace it immediately. If a needle puncture should occur, notify the lab manager or pro
10、ject coordinator immediately so that appropriate actions may be taken. ACTN Laboratory Technologist Committee Version 1.1 ACTN Dried Blood Spots Procedure 11 March 2009 3 4. SPECIMEN COLLECTION 4.1 Heel stick Procedure The Unistick 2 device has a penetration depth of 1.8 mm and a convex tip for accu
11、rate positioning that was designed to eliminate the risk of accidental needle stick injury or cross-infection as well as reduce the risk of osteomyelitis of the heel. Preferably, the baby should be in a supine position with the knee at the edge of the table or the baby may be held in the mothers arm
12、s. This position allows for the foot to hang lower than the torso, improving blood flow. Prepare the Unistick 2 device by depressing the pink plunger until it clicks. Twist the plunger until it breaks off and remove it from the device. The Unistick 2 device is now ready to trigger. When the baby is
13、in an acceptable position for this procedure, clean the incision area of the heel with an alcohol pad and allow the heel to air dry. Do not touch the incision site or allow the heel to come into contact with any non-sterile item or surface. Wrap your index finger around the base of the heel and your
14、 thumb around the ankle. See figure 1.0 below The Unistick 2 heel-stick device is properly positioned against the heel and triggered. The first drop of blood is wiped away using a dry sterile gauze pad. The blood is then allowed to flow to be collected on the dried blood spot card. To do this, gentl
15、y touch the filter paper card to the blood spot. Allow the card to absorb the blood until the circle is full. You should be able to obtain 5 spots of blood on a card. You may need to squeeze the heel to obtain more blood; however, do not milk the heel as it mixes interstitial fluid with the blood. O
16、nce all 5 spots are obtained, gently press a sterile pad to the incision site until bleeding has ceased. The babys heel must then be monitored for late bleeding and inflammation. Placing a sterile pad over the wound will help prevent a hematoma from forming. Note: Bandaging the babys foot is a contr
17、oversial issue because of skin sensitivity and potential bandage aspiration. However, the incision should be noted by the primary care nurse to ensure that the heel can be monitored for bleeding and inflammation. A bandage is not necessary as long as the bleeding has stopped before the child leaves
18、the clinic. Hold the Protein Saver 903 Card without touching the filter paper (See figure 2 below). Care should be taken to not touch the DBS circle once blood is applied. Allow the blood spot to air dry, with out card flap over spots, in a clean dry place (see drying racks Figure 3) that is protect
19、ed from rodents or insects and direct sunlight for at least 4 hours (drying overnight may be necessary in areas with higher humidity). Place ACTN Laboratory Technologist Committee Version 1.1 ACTN Dried Blood Spots Procedure 11 March 2009 4 the card in a gas impermeable bag with a desiccant pack and
20、 humidity indicator card. (See Figure 4). Note: Do not use electric dryer or oven to dry cards. Store no more than one card per bag. Do not heat, stack or allow DBS to touch other surfaces during the drying process. Once in the bag spots can be stored at room temperature, about 15-30o C or - 20 to -
21、70 o C according to protocol MOPS and/or LPC until shipped to the processing or receiving laboratory. Figure 1. Heel stick Placement and Device Figure 2. Holding the Protein Saver 903 Card without touching the filter paper Field Cod ACTN Laboratory Technologist Committee Version 1.1 ACTN Dried Blood
22、 Spots Procedure 11 March 2009 5 Figure 3. Whatman Protein Saver 903 Card Drying Racks and Common drying rack ACTN Laboratory Technologist Committee Version 1.1 ACTN Dried Blood Spots Procedure 11 March 2009 6 Figure 4. DBS Card in a gas impermeable bag with a desiccant pack and humidity indicator c
23、ard ACTN Laboratory Technologist Committee Version 1.1 ACTN Dried Blood Spots Procedure 11 March 2009 7 4.2 Collection of Dried Blood Spot from Finger Stick 4.2.1 Clearly label the Protein saver 903 Card with Patient ID and Date, Protocol, Visit ID. 4.2.2 Disinfect selected site and prick using lanc
24、et/needle. 4.2.3 Uniformly saturate entire circle by quickly and gently touching NOT PRESSING the puncture site to the filter paper. Dont touch the Whatman Protein saver 903 Card at any stage of collection. See examples of good and bad spots Section 7.0 4.2.4 After collecting 5 dried blood spots, cl
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