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Medisch Labo Bruyland

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Abstract bachelorproef 2016-2017The detection of Extractable Nuclear Antibodies with two methods 

The aim of this study is to compare two devices (EuroBlotMaster and Phadia 250) for analysing antinuclear antibodies.

Antinuclear antibodies are autoantibodies that are used in the diagnosis of systemic autoimmune diseases such as Lupus Erythematosus, Sjögren’s syndrome, Systemic sclerosis, Mixed connective tissue disease, Polymyositis and Dermatomyositis. Examples of autoantibodies are anti-dsDNA, ant-Ro, anti-La, anti-Sm, anti-RNP, anti-Scl etc. This autoantibodies  are also called antinuclear antibodies.

The first step is a screening with Hep-2000® cells. The result is a fluorescent pattern and a titer. If the titer is at least 1/80, additional tests are done. This can be done on the EuroBlotMaster or/an Phadia 250.

The Phadia 250 uses the Fluorescence Enzyme immunoassay (FEIA) technique. First there is a screening with the Symphony. If this is positive, all the autoantibodies are separately determined. Autoantibodies that are tested; anti-dsDNA, anti-CENP, anti-Jo1, anti-Scl-70, anti-Sm, anti-RNP, anti-Ro52/60, anti-La and anti-RNP-70.

The EuroBlotMaster with the immunoblot technique, uses strips which are stained by indirect immunostaining. From this, a band pattern is observed by using the EuroLineScan. This scan converts an intensity to a qualitative observation. This strip can detect sixteen autoantibodies.

There is a good agreement between the two methods for most of the tested antibodies. Another advantage of the EuroBlotMaster is the detection of six other antibodies, that can help in the further diagnostic of auto-immune diseases. The workflow for ENA detection will be changed. After a screening with Symphony on Phadia 250, positive results are further analysed with the immunoblot technique with the EuroBlotMaster.

Generally it can be decided that the results on the EuroBlotMaster and the results on the Phadia 250 are comparable. 

Samenvatting eindwerk 2014-2015: Verification (implementation validation) of a HPLC analyser for the measurement of HbA1c
Haemoglobin is a protein in the erythrocytes that caries oxygen to the tissues. When glucose binds to haemoglobin, haemoglobin A1c is formed.
The detection of HbA1c is useful in the diagnosis and evaluation of treatment in diabetics.
In this study the Menarini HA 8180V HPLC analyser for HbA1c is validated and compared with Tosoh G8. The principle of both analysers is reversed phase cation-exchange chromatography.
The validation/verification protocol includes verification of within-run, between-run, correlation and trueness. 1000 patient samples were tested and samples from extern quality controle 2014 were requested in the WIV (institute of health).
Within-run variation for low (42 mmol/mol), middle (61,1 mmol/mol) and high (88,7 mmol/mol) concentrations, are respectively 0%, 0,36% and 0,54%.
Between-run variation for level 3 (25,0 mmol/mol), 4 (33,0mmol/mol) and 5 (73,6 mmol/mol) concentrations, are respectively 0%, 0% an 1%.
Correlation between Tosoh G8 and Menarini HA 8180V is good, the correlation coefficient is 0,996.
The results of trueness, both analysers had for low IFCC levels and middle IFCC levels an aberration of 1, for high IFCC levels the Tosoh G8 had an aberration of 1 and the Menarini HA 8180V had an aberration of 0. The results for trueness are excellent according to the criteria of the WIV for both analysers.

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