Qualitative tests for globulins in spinal fluids

Pandy Test

The reagent consists of a saturated solution of pure, white-pink phenol in water. About 10 gm. of phenol (free of any brown-black coloration) is suspended in 100 ml. of water and the mixture gently warmed with shaking at 50 degree to 60 degree of Celcius. The material is transferred to a wide-mouth brown bottle and allowed to settle at room temperature (25 degree plus minus 2 degree of Celcius) for 48 to 72 hours before use. The reagent must be clear, colorless, and nonturbid. It is stored at room temperature in a brown bottle away from direct light and away from any cooling drafts.

Procedure

To 1.0 ml. of Pandy reagent in a 10 x 75 mm. test tube, add 0.10 ml. of clear CSF fluid. Mix the tube by inversion and allow to stand for exactly 3 minutes, and then examine it for the presence of any opalescence or turbidity by viewing it against a black background. The turbidity formed indicates the presence of greater than normal amounts of globulins, primarily gamma globulins. Grade the degree of turbidity from negative to ++++. The observed turbidity is scored similarly to the scheme described in Table 5.2 for the heat test for urine protein, but the protein equivalents are different. No consistent relationship between turbidity score or appearance and globulin concentration has been established. A Pandy reading of trace to 1+ will reflect a total protein concentration of 50 to 80 mg./100 ml., with a globulin level of about 20 to 40 mg./100 ml. For a ++ (2+) reading, the respective values are 100 to 160 and 35 to 70 mg./100 ml. In many cases an increase in albumin is accompanied by a proportionate increase in the globulins. In other cases (e.g. poliomyelitis) there is a disproportionate increase in the albumin fraction, giving cause to an increase in total protein without a corresponding Pandy reaction, In multiple sclerosis, in which only gamma globulins are increased. Pandy readings of 1+ or 2+ are obtained even though the total protein is within the normal range.