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:• 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

Flowchart of process

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

The equipment for specimen collection

13

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

Labelling of storage tubes

Transfer of storage tubes into freezer

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

• 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.

Safety issues

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