Laboratory of Immunology

Director - Assistant Professor: L. Skoura

Stuff
Senior Consultant and Director of Immunology Department
V. Tsavdaridou-Fola, Medical Biopathologist, PhD
+302310993545, +302310993542, vaso.tsavdaridou@gmail.com

Junior Consultant
E. Gounari, Medical Biopathologist , MSc, PhD
evigounari@yahoo.com

Scientist
É. Æoumpourtikoudi

Ôechnicians
Å. Koulouri
F. Faka
C. Êandilidou
G. Makrovasili



Laboratory location and working hours
The immunology department is located on the underground floor of the central part of AHEPA University Hospital, together with the rest of the Central Laboratory Departments.

The working hours are Monday-Friday 8am-3pm. There is no weekend or holiday laboratory on-call service, but samples can be delivered to medical staff on duty.

Overview and core objectives
We have supported immunology testing of hospitalized patients and outpatients for almost 25 years, as well as scientific and educational activities. Our primary objective is to offer the best possible medical services to patients regarding immunological testing, by aiming at quality, interdisciplinary collaboration and continuous training of medical, scientific and technological staff.

Tests performed and results

Tests performed

Laboratory methods used
The tests performed include determination of proteins in biological fluids, identification of autoantibodies, measurement of specific IgE antibodies (RAST), detection of IgG oligoclonal bands in CSF, and immunophenotyping. For this purpose we use a variety of laboratory methods consisting of indirect immunofluorescence, ELISA, rate nephelometry, immunoluminometry, capillary electrophoresis and immunotyping, isoelectric focusing, and flow cytometry.  

Results
Medical staff evaluates the results through informatics system used in the hospital (LIS). Whenever appropriate, clinical comments regarding interpretation are made on the report. The reference range for each test is included in the report in order to assist interpretation. Abnormal results are reported and may be phoned, if appropriate. It is important that as much relevant clinical information as possible is included in the request, in order to aid proper evaluation.

Please do not require profiles or screens, but specify the appropriate tests. If in doubt,  please contact with the laboratory staff.

Medical staff is always pleased to answer queries regarding the choice of the appropriate laboratory method and the interpretation of tests.

Results are given to patients by their attending clinicians and to outpatients by the staff of outpatient clinics. For patient samples referred from other hospitals or   people with difficulties in access to the hospital, results are sent by mail or FAX.

Categories of tests performed according to method and/or clinical significance

  • Immunophenotyping of peripheral blood, bone marrow, CSF and other biological fluids during diagnostic investigation or monitoring of hematologic neoplasms, autoimmune diseases, primary or secondary immunodeficiencies and infections.

Type of test

Laboratory method

Acute leukemia immunophenotyping

Flow cytometry

Diagnostic investigation of lymphoproliferative diseases

Flow cytometry

Diagnostic investigation of multiple myeloma

Flow cytometry

Determination of CD4+, CD8+T cells’ percentage and CD4/CD8 ratio

Flow cytometry

Determination of lymphocytic subsets

Flow cytometry



  • Quantitative determination of acute phase proteins-markers of inflammation

Type of test

Laboratory method

CRP

nephelometry

CRP-hs

nephelometry

procalcitonin

immunoluminometry

 



  • Quantitative determination of proteins of variable clinical significance (markers of inflammation, haemolysis, etc)

Type of test

Laboratory method

Ceruloplasmin  

 

nephelometry

  A1-antithrypsin

 

nephelometry

  Haptoglobin

 

nephelometry

  Soluble and intracellular cytokines

 

ELISA, flow cytometry for research only



  • Investigation of monoclonal gammopathies

Type of test

Laboratory method

Quantitative determination of IgA,  IgG, IgM, IgE serum immunoglobulins

nephelometry

Quantitative determination of kappa and lambda serum and urine light chains

nephelometry

Quantitative determination of free kappa and lambda serum light chains (free ê/ë ratio)

nephelometry

Quantitative determination of  â2 microglobulin 

nephelometry

Detection of monoclonal protein(s) in serum  (qualitative)

Capillary zone serum electrophoresis

 

Characterization of the type of serum monoclonal protein(s) (qualitative)

 

Capillary zone immunotyping &/or immunofixation with appropriate antisera



  • Investigation of demyelinating diseases of central nervous system

Type of test

Laboratory method

Detection of  IgG oligoclonal bands in CSF

Isoelectric focusing and immunofixation

Quantitative determination of  IgG immunoglobulin in CSF  

nephelometry

Quantitative determination of  IgG immunoglobulin in serum

nephelometry

Quantitative determination of  albumin in CSF

nephelometry

Quantitative determination of  albumin in serum

nephelometry

Determination of IgG INDEX  

 

 



  • Investigation of complement-related disorders (e.g. immunodeficiencies, autoimmunity)

Type of test

Laboratory method

Quantitative determination of C3, C4 complement components

 

nephelometry

Quantitative determination of total complement haemolytic activity (CH50)

 

ELISA

Quantitative determination of c1 inhibitor

 

ELISA

Quantitative determination of circulating immunocomplexes(CIC)

 

ELISA



  • Investigation of autoimmune disorders by autoantibody detection

Type of test

Laboratory method

 Antinuclear antibodies (ÁÍÁ)

 

Indirect immunofluorescence (IIF)

Anti-ds DNA antibodies (anti-dsDNA)

 

Indirect immunofluorescence (IIF)

Antibodies to extractable nuclear antigens (ENAs SCREEN) 

 

ELISA

 

   Antibodies to extractable nuclear antigens (anti-Sm)

 

ELISA

   Antibodies to extractable nuclear antigens (anti-Rï/SS-A)

 

ELISA

   Antibodies to extractable nuclear antigens (anti-Lá/SS-B)

 

ELISA

   Antibodies to extractable nuclear antigens (anti-Scl-70)

 

ELISA

   Antibodies to extractable nuclear antigens (anti-Sm /RNP)

 

ELISA

   Antibodies to extractable nuclear antigens (anti-Jo-1)

 

ELISA

Anticardiolipin antibodies (ACL – IgG & IgM)

 

ELISA

Anti-â2-glycoprotein I antibodies (anti-â2-GPI – IgG & IgM)

 

ELISA

Anti-neutrophil cytoplasmic antibodies (ANCA – anti-MPO & anti-PR3)

 

ELISA

Anti-glomerular basement membrane antibodies (anti-GBM)

 

Dot-blot assay

Anti-cyclic citrullinated peptide antibody (anti-CCP)

 

ELISA

Anti-mitochondrial antibodies (AMA)

 

Indirect immunofluorescence (IIF)

Anti-smooth muscle antibodies (ASMA)

 

Indirect immunofluorescence (IIF)

Anti-gastric parietal cell antibodies (APCA)

 

Indirect immunofluorescence (IIF)

Anti-Liver Kidney Microsomal Antibodies (anti-LKM1)

 

Indirect immunofluorescence (IIF)

Anti-Endomysial Antibodies (EMA)

 

Indirect immunofluorescence (IIF)

Tissue transglutaminase antibody (anti-tTG – IgG & IgA

 

ELISA

Anti-Gliadin Antibodies (AGA - IgG & IgA

 

ELISA



  • Investigation of allergic disorders by detection of allergen-specific IgE antibodies

Type of test

Laboratory method

  • Quantitative determination of specific IgE antibodies against inhaled allergens

 

ELISA

  • Quantitative determination of specific IgE antibodies against food allergens

 

ELISA

  • Quantitative determination of specific IgE antibodies against drug allergens

 

ELISA



Requesting tests and sample delivery
Ordering tests

A request form must accompany all specimens sent to the laboratory. The form must clearly state the following information for unequivocal identification of patient and specimen as well as for the reason of ordering:

  • Patient full name
  • Requesting doctor' s full name
  • Full name of the person collecting the sample
  • Referring department and hospital
  • Age
  • Sex
  • Specimen type
  • Date and time of specimen taken
  • Tests required
  • All relevant clinical details

The requesting doctor must sign the request form and write his full name to enable contact for information and clinically significant results.
Hazardous samples must be clearly identified.

Specimens

  • specimen labelling

The specimens must be labelled with the patient details and date of collection, which must agree with the respective information of the requesting form. It is the responsibility of the person collecting the sample to ensure that it is correctly labelled. It is not possible to change the patient's details once the sample has been sent to the laboratory. Specimens not properly or clearly labeled will be discarded.

  • type of specimen and specimen collection

Whole peripheral blood and bone marrow: use tube with EDTA anticoagulant (take care for correct mixing) for flow cytometric immunophenotyping.
Serum (clotted peripheral blood): use tube without anticoagulant for the performance of tests other than immunophenotyping in peripheral blood.
Biological fluids: use tube with heparin anticoagulant for flow cytometric immunophenotyping and/or tube without anticoagulant for all other tests.

  • quality and quantity of specimens

Freshly drawn samples from fasting individuals are preferred. Samples must be adequate in quantity for the determination of all tests required. Haemolytic and/or lipaemic samples are rejected.
Approximately 1 mL of CSF and serum respectively are needed for investigation of demyelinating disease of CNS (i.e. isoelectric focusing/immunofixation and IgG index determination).
One vial of EDTA anticoagulated whole blood is usually sufficient for immunophenotyping.

  • sample packaging and transport

For samples sent from an external laboratory or clinical department, it is the responsibility of the requesting laboratory or clinic to ensure that the samples are correctly packed, labelled, accompanied with the appropriate paperwork and transported, so that public health and validity of results are ensured. Samples must be sent at 2-8oC.

  • sample delivery and tests' performance

Samples (preferably from fasting individuals) must be delivered after phone arrangement from Monday to Friday, during early morning working hours (8-11 a.m.). If tests are required urgently, please contact the laboratory and discuss with medical staff.
Samples for flow cytometric immunophenotyping must be delivered after phone arrangement from Monday to Thursday, during early morning working hours (8-11 a.m.). For urgent cases, please contact with the laboratory before sample delivery.

Guidelines for Receiving and Handling of Samples for the Immunology Laboratory

Test

Sample
type

Tube type

Necessary quantity

Patient preparation

Sample delivery time

Laboratory report

Notes

Capillary electrophoresis, immunosubtraction and immunofixation of serum proteins

whole blood

Without
anticoagulant

4 ml

No

< 6 h

8-10
days

 

IgG,  IgA,  IgM serum immunoglobulins

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

2-5 days

 

kappa and lambda serum light chains

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

2-5 days

 

Free kappa and lambda serum light chains

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

5-7 days

 

IgÅ serum immunoglobulin

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

2-5 days

 

IgG index and detection of IgG oligoclonal bands in CSF

whole blood
&
CSF

Without
anticoagulant

4 ml of blood &
at least
0,5 ml of CSF
received at the same day

fasting

< 6 h

25-30 days

 

Complement factor C3

whole blood

Without
anticoagulant

4 ml

fasting

< 3 h

2-5 days

 

Complement factor C4

whole blood

Without
anticoagulant

4 ml

fasting

< 3 h

2-5  days

 

Rheumatoid factor

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

2-5  days

 

 

Guidelines for Receiving and Handling of Samples for the Immunology Laboratory

Test

Sample
type

Tube type

Necessary quantity

Patient preparation

Sample delivery time

Laboratory report

Notes

CRP (C-Reactive Protein)

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

1-3 days

 

HPT (Haptoglobin)

whole blood

Without
anticoagulant

4 ml

No

< 6 h

2-5 days

 

CER (Ceruloplasmin)

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

2-5 days

 

AAT (Álpha1-Antitrypsin)

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

2-5 days

 

ASTO (Antistreptolysin O)

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

2-5  days

 

 Â2M (Â2 Microglobulin)

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

5-7  days

 

CRP hs (high sensitivity C-Reactive Protein)

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

5-7 days

 

PCT (Procalsitonin)

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

1-3  days

 

 

 

 

 

 

Guidelines for Receiving and Handling of Samples for the Immunology Laboratory

Test

Sample
type

Tube type

Necessary quantity

Patient preparation

Sample delivery time

Laboratory report

Notes

Ô-Lymphocytes'
surface antigens
(AIDS)

whole blood

Ê3 EDTA

2 ml
(1 complete blood count tube)

No

< 3 h

3-5 days

Sample delivery Tuesday-Thursday

Determination
of lymphocytic subpopulations

whole blood
bone marrow
CSF
ascetic fluid
pleural fluid

Ê3 EDTA
or heparin

2 ml whole blood or bone marrow
(1 complete blood count tube)/at least
0,5 ml of other fluids

No

< 6 h

15 days

Sample delivery by appointment

Acute leukaemia Immunophenotyping

whole blood
bone marrow
CSF
ascetic fluid
pleural fluid

Ê3 EDTA
or heparin

4 ml whole blood or bone marrow
(2 tubes)/at least
0,5 ml of other fluids

No

< 6 h

15 days

Prearrangement
with the Lab

Mature B-  or T- cell neoplasms immunophenotyping

whole blood
bone marrow
CSF
ascetic fluid
pleural fluid

Ê3 EDTA
or heparin

4 ml whole blood or bone marrow
(2 tubes)/at least
0,5 ml of other fluids

No

< 6 h

15 days

Sample delivery by appointment

T Cell Receptor (TCR) Vb repertoire determination

whole blood

Ê3 EDTA

2 ml
(1 tube)

No

< 6 h

15 days

Sample delivery by appointment

Plasma cell myeloma

Bone marrow

Ê3 EDTA

2 ml
(1 tube)

No

< 6 h

15 days

Sample delivery by appointment

 

 

Guidelines for Receiving and Handling of Samples for the Immunology Laboratory

Test

Sample
type

Tube type

Necessary quantity

Patient preparation

Sample delivery time

Laboratory report

Notes

Nuclear Autoantibodies (ÁÍÁ) -
Centromere pattern Nuclear Autoantibodies (ACA) included

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

5-7  days

 

dsDNA Autoantibodies

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

7-10  days

 

Mitochondrial Autoantibodies (ÁÌÁ)

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

7-10  days

 

Anti-smooth muscle Autoantibodies (ÁSMA)

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

7-10  days

 

Liver/Kidney Microsomal Autoantibodies (ÁÌÁ) (LKM1)

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

7-10  days

 

Parietal cell Autoantibodies (ÁPCA)

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

7-10  days

 

Endomysium Autoantibodies (EMA)

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

20-25  days

 

Tissue Transglutaminase Autoantibodies (Anti-tTG) IgG - IgA

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

20-25  days

 

Gliadin Autoantibodies
(AGA) IgG - IgA

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

20-25  days

 

Cyclic Citrullinated Peptide Autoantibodies (anti-CCP)

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

30  days

 

Glomerular Basement Membrane/Collagen IV-á3 Autoantibodies (anti-GBM)

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

1 - 3  days

Prearrangement
with the Lab

 

 

Guidelines for Receiving and Handling of Samples for the Immunology Laboratory

Test

Sample
type

Tube type

Necessary quantity

Patient preparation

Sample delivery time

Laboratory report

Notes

Extractable Nuclear Antigen Autoantibodies (ENA-6 SCREEN)

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

20-25  days

 

Sm-RNP Autoantibodies

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

20-25  days

 

Sm Autoantibodies

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

20-25  days

 

SS-A/Ro Autoantibodies

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

20-25  days

 

SS-B/La  Autoantibodies

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

20-25  days

 

Scl-70  Autoantibodies

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

20-25  days

 

Jo-1 Autoantibodies

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

20-25  days

 

Cardiolipin Autoantibodies (ACL)
IgG - IgM

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

20-25  days

 

B-2 Glycoprotein Autoantibodies (â-2 GPI)
IgG - IgM

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

20-25  days

 

Proteinase-3  Autoantibodies (PR3/c-ANCA)

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

20-25  days

 

Myeloperoxidase  Autoantibodies (MPO/p-ANCA)

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

20-25  days

 

 

 

 

 

Guidelines for Receiving and Handling of Samples for the Immunology Laboratory

Test

Sample
type

Tube type

Necessary quantity

Patient preparation

Sample delivery time

Laboratory report

Notes

Total classical pathway complement activity (CH50)

whole blood

Without
anticoagulant

4 ml

fasting

< 3 h

30 days

 

C1-Inhibitor (C1-INH)

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

30 days

 

  Circulating Immune Complexes (CIC)

whole blood

Without
anticoagulant

4 ml

fasting

< 6 h

30 days

 

 

 

The necessary quantity reported in the above tables regards individual tests performed from whole blood without anticoagulant. In case of more than one required tests, 0.5 ml of whole blood is needed per test, additionally.

For any further information, please contact the laboratory staff.


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