Detection of enzyme deficiencies in red blood cells

摘要

Simple rapid method duitable for the mass screening of blood samples to detect deficiencies of glucose-6-phosphate dehydrogenase (G-6-Pd) in red blood cells, is as follows:- Blood sample (1 pt.) is added to 10 pts. of an aqs. soln. made up of 1 pt. of a 0.002-0.3M pref. 0.01M aqs. soln. of glucose-6-phosphate, 1 pt. of a 0.001875-0.075 M pref. 0.0075M aqs. soln. of oxidised triphosphopyridine nucleotide (TPN), 2 pts. of an aqs. soln. of a red blood cells lysing agent (partic. 0.1 satd. to satd., pref. satd. soln. of digitonin or 0.1-20% pref. 1% soln. of saponin), 3 pts. of 0.025-1.25M pref. 0.25M K phosphate buffer at pH 6.5-8.0 pref. 7.4, and 3 pts. of H2O. Mixture is allowed to react, and then spotted onto filter paper. Dried spot is then irradiated with u.v. light of lambda = 340-370 nm; the spot only fluoresces if there is no deficiency of G-6-PD in the red blood cells of the blood sample. Details are given of a similar process for the detection of galactose-1-phosphate uridyl transferase deficiencies.