Appendix 5. Protein estimation

5.1 Pandy test

Pandy test is used to detect an increase of protein in the cerebrospinal fluid (CSF).
The normal range of protein in CSF is 0.20-0.45 g/litre.
The Pandy test is positive when protein is superior to 0.45 g/litre.

Equipment

– Gloves
– Pandy reagent
– Pasteur pipettes
– Conical centrifuge glass tube or test tube
– 1 ml pipettes

Preparation of 500 ml of Pandy reagent

Pandy is a saturated phenol solution.
– Weigh 30 g of phenol and transfer it in a 1000 ml bottle.
– Add 500 ml of distilled water and shake vigorously.
– Leave to stand for one day.
– Check whether any phenol remains undissolved:
• If so filter, the solution is ready.
• If all the phenol has dissolved, add a further 10 g of phenol and wait another day before filtering.

Pandy reagent is a highly corrosive and toxic solution:
– Label the bottle and mark it corrosive and poisonous.
– Wash hands after the preparation.

Technique

– Place 1 ml of Pandy reagent in a centrifuge tube.
– Add 3 drops of CSF, drop by drop.
– After each drop, look for a white cloud in the tube.
– In order to facilitate the reading, place a black surface behind the tube.

Results

– Presence of a white precipitate: Pandy test positive;
– Absence of a white precipitate: Pandy test negative.

5.2 Rivalta test

The Rivalta test is used to detect an increase of protein in the body fluid (pleural fluid, ascites).
The test is positive when the proteins are superior to 30 g/litre.

Equipment

– Gloves
– Rivalta reagent
– Pasteur pipettes
– Conical centrifuge glass tube or test tube
– 5 ml pipette

Preparation of 100 ml of Rivalta reagent

– Place 50 ml of distilled water in a 100 ml measuring cylinder.
– With a 5 ml pipette, add 3 ml of glacial acetic acid and make up to the 100 ml mark with the remaining 50 ml of distilled water.
– Transfer the solution in a bottle.

Technique

– Place 2 ml of Rivalta reagent in a centrifuge tube.
– Add 3 drops of pleural fluid/ascites, drop by drop.
– After each drop, look for a white cloud in the tube.
– In order to facilitate the reading, place a dark surface behind the tube.

Results

– Presence of a white precipitate: Rivalta test positive;
– Absence of a white precipitate: Rivalta test negative.