Introduction
- Every lab has its own organization.
- Our humoral lab belongst to hematology lab, while it performs several tests for clinical biochemistry.
- Mostly we used serum tube (on cellular part, EDTA), except for fibronectin and cryocrite (?)
Machines and its tests
Phadia ImmunoCap 250
- ELISA: antigen on the bottom --> serum from patients with antibodies --> conjugate --> serum
- 'vertical' wells
- Test for allergy (numbered: food birds), IgE (in clinical chemistry), IgG
- Test for an autoimmune testing (i.e. anti CCP, anti gliadin (IgG), anti PR3 and anti MPO (both are performed only when ANCA is positive).
Siemens BNII
- Nephelometer: measures the turbidity.
- protein in plasma (antigens) bind with antibodies in reagentia
- to measure protein in serum, i.e. free light chain, heavy chains (IgG and its subtypes), prealbumin (nutrition state especially for older patients), trasnferrin, hsCRP, haptoglobulin.
Sebia electrophoresis
- protein in serum through the gel electrophoresis will show bands. positive protein will go to negative pole (such as IgM and IgG; IgG travels the furtherst because it is small), negative will go to positve (such as prealbumine and albumine). Adding colour is needed to show the band pattern.
- high resolution: serum without adding any reagens.
How to read: any bands in beta or gamma?
- immunofixation: serum, addition of anti IgG or anti IgA, or anti IgM.
How to read: look at the Ig G or IgA or IgM, look at the same height any kappa or lambda? for example band on IgG and at the same height band on kappa: IgG kappa. If there is another band in light chain than it must be free light chain also present.
- Bence Jones protein: free kappa and/ or lambda light chain in urine.
Zenit up
- to be honest, this machine makes a lot of noise.
- it incubates the serum with for example ANA or ANCA on tissues (available from manufactures such as Biorad). The tissues can be skin, stomach (parietal cells) kidney liver, glomerular basal membrane mostly to detect automimmune disease. The tissues are originated from rat, monkey or human.
- The fluorescense can be then observed by using fluorescense microscope.
- When ANA is positive (anti dsDNA and anti ENA should be performed), when ANCA is positive (anti MPO and PR3 can be performed).
Others
- Viscosity test using a cappilary column to determine hoe 'thick' the blood. The blood can be 'thick' due to disease such as multiple myeloma.
- Complement agglutination test.
- To test the concentration of serum that destroy 50% of red blood cells. Standard curve is made by putting serum concentration on the x as and the clarity on y-as.
- To do this tests, red blood cells from sheep are mixed with IgG anti sheep. Than serum is added. The reaction should be stopped directly in cold water.
- Important to have the serum at 4C. Otherwise the results will be underestimation.
- Every lab has its own organization.
- Our humoral lab belongst to hematology lab, while it performs several tests for clinical biochemistry.
- Mostly we used serum tube (on cellular part, EDTA), except for fibronectin and cryocrite (?)
Machines and its tests
Phadia ImmunoCap 250
- ELISA: antigen on the bottom --> serum from patients with antibodies --> conjugate --> serum
- 'vertical' wells
- Test for allergy (numbered: food birds), IgE (in clinical chemistry), IgG
- Test for an autoimmune testing (i.e. anti CCP, anti gliadin (IgG), anti PR3 and anti MPO (both are performed only when ANCA is positive).
Siemens BNII
- Nephelometer: measures the turbidity.
- protein in plasma (antigens) bind with antibodies in reagentia
- to measure protein in serum, i.e. free light chain, heavy chains (IgG and its subtypes), prealbumin (nutrition state especially for older patients), trasnferrin, hsCRP, haptoglobulin.
Sebia electrophoresis
- protein in serum through the gel electrophoresis will show bands. positive protein will go to negative pole (such as IgM and IgG; IgG travels the furtherst because it is small), negative will go to positve (such as prealbumine and albumine). Adding colour is needed to show the band pattern.
- high resolution: serum without adding any reagens.
How to read: any bands in beta or gamma?
- immunofixation: serum, addition of anti IgG or anti IgA, or anti IgM.
How to read: look at the Ig G or IgA or IgM, look at the same height any kappa or lambda? for example band on IgG and at the same height band on kappa: IgG kappa. If there is another band in light chain than it must be free light chain also present.
- Bence Jones protein: free kappa and/ or lambda light chain in urine.
Zenit up
- to be honest, this machine makes a lot of noise.
- it incubates the serum with for example ANA or ANCA on tissues (available from manufactures such as Biorad). The tissues can be skin, stomach (parietal cells) kidney liver, glomerular basal membrane mostly to detect automimmune disease. The tissues are originated from rat, monkey or human.
- The fluorescense can be then observed by using fluorescense microscope.
- When ANA is positive (anti dsDNA and anti ENA should be performed), when ANCA is positive (anti MPO and PR3 can be performed).
Others
- Viscosity test using a cappilary column to determine hoe 'thick' the blood. The blood can be 'thick' due to disease such as multiple myeloma.
- Complement agglutination test.
- To test the concentration of serum that destroy 50% of red blood cells. Standard curve is made by putting serum concentration on the x as and the clarity on y-as.
- To do this tests, red blood cells from sheep are mixed with IgG anti sheep. Than serum is added. The reaction should be stopped directly in cold water.
- Important to have the serum at 4C. Otherwise the results will be underestimation.
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