Name :Dorene Chen


Hello people, it's my turn to write blog again.

To date, I have been to biochemistry (done blog), haematology and serology department.

Serology


The tests I will be going through are:


1)AMC (Anti microsomal/thyroid peroxidase antibodies = same thing but different name)
2)ATG (Antibodies to ThyroGlobulin).

3)ANF slide (Anti nuclear Factor)


AMC/ ATG testing ( methods are the same so i join them together =D)
AMC/ATG testing tests for presence of autoimmune antibodies. It uses gelatin particles sensitized with human Thyroglobulin (Tg) or thyroid peroxidase (TPO) antigen for agglutination. serum containing specific antibodies will react with gelatin particle to form a smooth mat of agglutinated particles in the microtitration plate (aka "ELISA" plate) testing tests for

Autoimmune thyroid disease is autoimmune disease of the thyroid gland in which the patient's own immune system attacks his own thyroid. In patients suffering from this disease may secrete anti thyroglobulin and Anti microsomal antibodies against their own thyroid antigen (thyroglobulin, microsomal/thyroid peroxidase antigen are found in thyroid).


Thyroglobulin (Tg) is a protein found and produced by thyroid and T3, T4 are produced from this protein. T3, T4 are hormones that aids in transporting protein, controlling metabolism and they affect almost every process in the body.

Microsomal/ thyroid peroxidase (TPO) is an enzyme in thyroid that aids in the function of producing T3, T4 through oxidising iodide compounds in blood and to incorporate the oxidized iodide compounds into T3, T4.

Method (uses multi-well/microtitration plate aka "ELISA" plate):






Note: the picture above shows only the reagents to add in one row.
1)add 50µL of sample diulent to 1st well. Add 75 µL to 2nd-6th well.
2) add 10µL of positive control to first well. add 10 µL of sample to other row in the multiwell plate.
3) Do serial dilution by taking 25µL of suspension from 1st well into 2nd well. pipette and mix well. From 2nd well, pipette another 25 µL to 3rd. repeat until 6th well. discard away the 25µL from the 6th well.
4 ) Add 25µL unsensitized particles ( these particles will not cause any reaction, act as control) only to the 2nd well of the control.
5) Add 25 µL sensitized particles (these particles cause agglutination when antibodies is present) to the other well.
6) Incubate for 3 hours.

Positive result: Form large ring (dark purple color background with light purple ring shape in the center cause by agglutination). For very strong reaction, the dark purple color background disappears completely.

Negative result: Button shaped (Purple color throughout)