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应用巴比妥缓冲液琼脂糖凝胶电泳法测定血清LDH同工酶常出现LDH_5区带向负极移动,落在原点之后,同时LDH_(4-5)区带含量常偏低。为了纠正上述缺点,我们根据日本长岭光隆等提出的焦磷酸缓冲液琼脂糖电泳法进行测定与方法学探讨。我们对100例正常血清标本用本法与巴比妥法同时测定LDH同工酶的百分含量,本法测定结果;LDH_1 26.5±4.3,LDH_2 33.1±3.7,LDH_3 22.3±3.5,LDH_4 12.0±2.8,LDH_5 6.1±2.9,而巴比妥法测定结果:LDH_1 29.6±4.5,LDH_2 35.0±5.8,LDH_3 21.5±4.4,LDH_4 9.3±4.1,LDH_5 4.6±2.9。比较二法测定结果表明,本法对LDH_4和LDH_5区带较为灵敏,p值分别为P<0.05和P<0.01,有显著性差异,其次,本法在一定条件下,具有使电泳各区带推向沟前的良好效果,这是本法优於巴比妥法之处。在方法学上,我们对使用载体、冷却装置及固定液、洗脱等步骤均进行某些改进,使本法更为简便、更适合于临床应用,且便于应用光密度扫描。
Serum LDH isozymes were detected by barbiturate buffer agarose gel electrophoresis. LDH_5 band shifted to the negative pole, and the LDH_ (4-5) band content was often low. In order to correct the above shortcomings, we conducted the determination and methodological study according to the pyrophosphate buffer agarose electrophoresis proposed by Japan’s Changling Guanglong et al. 100 cases of normal serum samples by this method and barbiturate method simultaneously determine the percentage of LDH isoenzymes, the results of this method; LDH_1 26.5 ± 4.3, LDH_2 33.1 ± 3.7, LDH_3 22.3 ± 3.5, LDH_4 12.0 ± 2.8 , LDH_5 6.1 ± 2.9, and the results of barbiturate determination: LDH_1 29.6 ± 4.5, LDH_2 35.0 ± 5.8, LDH_3 21.5 ± 4.4, LDH_4 9.3 ± 4.1, LDH_5 4.6 ± 2.9. Compared with the results of two methods, the results showed that this method is sensitive to the LDH 4 and LDH 5 bands, with p values of P <0.05 and P <0.01, respectively. There is a significant difference between the two methods. The good effect that precedes the ditch is that this law is superior to the law of Babbitt. In methodology, we have made some improvements to the procedure of using carriers, cooling devices, fixatives and elution to make this method easier and more suitable for clinical applications and to facilitate the application of optical density scanning.