NDPK-A诊断试剂盒的研制

Establishment of NDPK-A diagnostic kit

目的建立一种灵敏、特异检测血清中二磷酸核苷激酶（NDPK-A）含量的酶联免疫吸附试验（ELISA）法和试剂盒,分析血清中的NDPK-A含量与血液病之间的关系。方法筛选NDPK-A多株单克隆抗体和多克隆抗体,以建立检测NDPK-A的双抗体夹心ELISA方法。测定了84例健康人,35例白血病与淋巴瘤等血液病患者血清的NDPK-A含量。结果单抗-多抗夹心ELISA法可测最适范围为1.56～100μg/L,回归方程为y=-0.0005x2＋0.0178x＋0.0892,相关系数r2=0.9993（y为A490nm,x为[NDPK-A]）,批内变异系数4.75%～6.32%,批间变异系数11.06%～13.84%,回收率为90.52%～105.16%。84例健康人血清NDPK-A含量为1.75±0.92μg/L,35例白血病与淋巴瘤患者血清的阳性检出率为16例。急性白血病,尤其是急性单核细胞白血病（M5）和非霍奇金淋巴瘤（NHL）患者血清中NDPK-A水平与健康对照差异较大,分别为12.97±9.73、20.00±18.55μg/L。结论建立了一种可应用于临床检测NDPK-A的ELISA方法,初步展示了NDPK-A含量检测的临床意义,NDPK-A含量检测可发展为白血病的辅助诊断和疗效判断指标。

Objective To establish a sensitive and a specific ELISA kit for detecting and analysing serum ND- PK-A and to explore the relationship between serum NDPK-A content and hemopathy. Methods The double-antibodies sandwich ELISA was established for detecting NDPK-A by self-made monoclonal antibody （mAb） and polyclonal antibody （pAb） against NDPK-A. In combination with the determination of NDPK-A, the investigation on the serum level of NDPK-A in 84 normal adults and 35 patients with leukemia or lymphoma was performed. Results The results showed that the optimal range of NDPK-A was 1. 56 - 100μg/L, the regression equation was y ： -0. 000 5x^2 ＋0. 017 8x＋0. 089 2 （r^2=0. 999 3）. The variation coefficient of intra-assay and inter-assay was 4.75 % -6.32% and 11.06%-13.84%, respectively. The recovery rate was 90.52%-105.16%. The mean level of serum NDPK-A level in 84 normal adults was 1.75±0.92 μg/L. The serum level of NDPK-A of 16 patients with leukemia or lymphoma was significantly higher than that of the healthy adults, especially in patients with AML-M5 or NHL, in which the level was 12.97±9.73 μg/L and 20.00±18.55 μg/L, respectively. Conclusion The sandwich ELISA established can be applied to measuring the serum level of NDPK-A, and might be developed into a useful tool to study the relationship between serum NDPK-A levels in leukemia and lymphoma or prognosis of these diseases. It would be important for planning an appropriate treatment strategy.