Tarkkanen P, Komor S, Cornely C, Hacke W, Greiling H
Clin Chem. 1979 Sep;25(9):1644-8.
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.
我们研究了生物发光法在测定血清和脑脊液中极低浓度脑型肌酸激酶亚基(CK-BB)时的灵敏度。为优化CK同工酶检测的灵敏度并消除可能的误差来源,我们使用抑制性抗-MM抗体以及沉淀性抗-MM和抗-BB抗体分离同工酶组分。生物发光检测结果与分光光度法值相关,总CK活性的r = 0.97,CK-B活性的r = 0.98。本方法的重现性与分光光度法相当,在低酶活性时甚至更好。在2 U/L测定总CK活性时,批内精密度对应的变异系数(CV)为9%;在13 U/L时,CV为5.8%。所有检测均在25摄氏度下进行。即使在这个低温下,低至0.2 U/L的CK活性也能被测定。在8例无任何脑细胞损伤证据的患者中,脑脊液中的总CK活性为x = 1.05±0.6 U/L,CK-BB活性为x = 0.7±0.4 U/L。这些患者血清中的CK-BB活性为x = 0.6±0.5 U/L。文中讨论了4例有短暂性或进行性脑细胞损伤患者的CK和CK-BB活性差异。