Bioluminescence assay of creatine kinase and its isoenzymes in serum and cerebrospinal fluid.
We examined the sensitivity of bioluminescence for the determination of very low concentrations of creatine kinase brain-type subunit (CK-BB) in serum and in cerebrospinal fluid. To optimize the sensitivity of CK-isoenzyme assays and eliminate possible sources of error, we separated the isoenzyme fractions by using inhibiting anti-MM and precipitating anti-MM and anti-BB antibodies. The results with the bioluminescence assay correlated with spectrophotometric values such that r = 0.97 for the total CK activity and r = 0.98 for the CK-B activity. The reproducibility of the present method was comparable with the spectrophotometric method and was even better at low enzyme activities. The within-series precision for assay of total CK activity at 2 U/L corresponded to a CV of 9%; at 13 U/L the CV was 5.8%. All the assays were carried out at 25 degrees C. Even at this low temperature, CK activities as low as 0.2 U/L could be determined. In eight patients without any evidence of cerebral cell damage, total CK activity in cerebrospinal fluid was x = 1.05 +/- 0.6 U/L, and CK-BB activity was x = 0.7 +/- 0.4 U/L. In sera of these patients CK-BB activity was x = 0.6 +/- 0.5 U/L. Differences in CK and CK-BB activities in four patients with transient or progressive brain-cell damage are discussed.
D001322:Autoanalysis; D003402:Creatine Kinase / Q000032:analysis* / Q000097:blood / Q000134:cerebrospinal fluid; D005459:Fluorides / Q000494:pharmacology; D006801:Humans; D007527:Isoenzymes / Q000032:analysis* / Q000097:blood / Q000134:cerebrospinal fluid; D007700:Kinetics; D008163:Luminescent Measurements; D008722:Methods; D013053:Spectrophotometry / Q000379:methods
D007527:Isoenzymes; D003402:Creatine Kinase; D005459:Fluorides