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Training Laboratory procedures. Georg Alfthan and Laura Lund. Helsinki 20-23.9.2010. Aim of training. The purpose of a European health survey is to collect data and biological samples from a nationally representative random population. The main goal is the core measurements:
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TrainingLaboratory procedures Georg Alfthan and Laura Lund Helsinki 20-23.9.2010
Aim of training • The purpose of a European health survey is to collect data and biological samples from a nationally representative random population. • The main goal is the core measurements: • Serum cholesterol, HDL cholesterol and glucose • Additional measurements:HbA1c, DNA • Special attention is paid to preanalytical conditions more demanding than needed for the core measurements. • All serum and plasma aliquots as well as two whole blood tubes are frozen at site.
Critical pre-analytical issues of blood collection • Fasting - 8-14 hours necessary for glucose, triglycerides • Position of subject • sitting • 10 min rest • lying 8% increase in blood volume • Tourniquet - short duration • Season - spring vs. winter (cholesterol) • Physical exercise • heavy training dehydration
Equipment and materials (1) • Blood tubes • - 2 plain gel • - 1 F-citrate • - 3 EDTA • Blood collection • pooling tube • needles • adapter • tournique, etc • skin tape • disinfectant for skin, table surface • Tube racks for • - blood tubes • - cryotubes • - counterbalance centrifuge tubes • Waste containers • - biological • - needle • - risk • Disposable gloves • - latex • - nitrile
Equipment and materials (2) • Storage tubes • - 1.5 ml cryotubes • - 3 ml plastic tubes • Storage boxes for • - cryotubes • - plastic tubes • - small whole blood • - large whole blood • Labels for • - blood tubes, cryotubes, plastic tubes • - tube rack • - storage boxes • - participant form
Equipment and materials (3) • Centrifuge, swinging bucket • Pipet, 1.5 ml, tips • Pasteur pipets, plastic pipets • Roller mixer • Styrofoam box (to simulate freezer) • Dry ice • Protective gloves • Timer • Thermometer • EHES MANUAL
Waste disposal • Biological waste container: Waste of human origin-contaminated with blood is disposed of according to the local facility regulations. • Needle disposal box: Full and sealed • disposed according to local facility regulations or sent to the NHES Laboratory. • Biological risk waste (suspected HIV, hepatitis or other contagious disease) should be sent to the NHES Laboratory according to its guidelines.
Preparing for blood sampling • The blood tubes are arranged in the designated tube rack according to the sampling chart. • * One subject per tube rack! • * Label blood tubes after venipuncture is done • * Extra tubes
Site of blood drawing Tube rack with participant’s ID label
Blood sampling chart -20 ºC
Tube labels The box shows type of sample or form or questionnaire. The abbreviation of the survey name with country code. The secondary key=unique person ID number The bar code with its numerical value=primary key
Drawing of blood samples • A personal label sheet is reserved for each participant • The forms are labelled according to the instructions of the manual. • The laboratory form is for documenting all events related to blood sampling and processing. • Fasting for 8-14 hours requested • 10-15 min resting before venipuncture • Sitting position
Documentation of blood collection, Recording form • ID of phlebotomist • Date and time of phlebotomy • Time from last meal • Number of tubes collected • Posture, and reason if supine • etc
Sample handling after centrifugation • Immediately after centrifugation, the caps must be removed except from the last two EDTA tubes • Place the bar code labels properly. The code must be upright or else the reading with barcode reader is impossible
Correct labelling of storage tubes Plastic tube
Storage box labels The box label sheet contains 2 identical labels. One label is placed on the front of the box and the other on the lid. Both labels should be visible when the lid is in place
Safety issues • It is prohibited to eat or drink in the laboratory area! • Wearing protective clothing is not allowed outside the laboratory area, like in the recreation room or cafeteria. • Table surfaces are desinfected by wiping with a 70 % ethanol solution at the end of the day. Any surface or object which is contaminated with blood or other human specimen must be disinfected immediately. • Disposable gloves must be used throughout handling of blood samples! • The sampling needle is removed directly into the needlebox from the adapter/holder without touching the needle • If the subject is suspected/has informed of HIV, hepatitis or other contagious disease, the nurse should use nitrile rubber gloves. The stasis and adapter are disposed of in the biological waste container as well as the gloves.
Ergonometry • The correct posture and equipment are important issues for maintaining your long term health status. • Chair height • Organization of equipment • More info in (Appendix)
Storage of specimen • Short term storage = - 20°C - Core measurements • Long term storage = - 70°C - Additional measurements National HES Laboratory