两种人血管内皮生长因子免疫测定试剂盒的研制及其对早期肿瘤的诊断价值探讨

目的 研制人血管内皮生长因子（hVEGF）放射免疫测定（RIA）试剂盒和化学发光免疫测定（CLIA）试剂盒，探讨其在早期肺癌和结直肠癌诊断中的价值。 方法 以2种试剂盒的技术指标为参考依据建立检测方法，确定磁微粒包被抗体的工艺方法及125I和吖啶酯标记抗体的工艺方法，并评价2种试剂盒的分析灵敏度、精密度、回收率。通过检测临床癌症（早期肺癌和结直肠癌）患者和正常人血清样本中的hVEGF，评价2种试剂盒对早期肿瘤诊断的灵敏度和特异度。2组间的比较采用两独立样本t检验。 结果 磁微粒与包被抗体的最佳比例为1 ml磁微粒∶1 mg 抗体；125I与标记抗体的最佳比例为55.5 MBq 125I∶0.1 mg抗体，吖啶酯与标记抗体的最佳比例为25 μg吖啶酯∶0.2 mg抗体。RIA和CLIA试剂盒的分析灵敏度分别为17.6、9.2 pg/ml。在精密度方面，CLIA试剂盒的批内变异性略高于RIA试剂盒，而批间变异性略低于RIA试剂盒。RIA和CLIA试剂盒的平均回收率分别为103.28%和101.85%，说明CLIA试剂盒检测的准确率更高。在临床样本检测方面，2种试剂盒对早期肺癌和结直肠癌的临床诊断灵敏度和特异度均可达90%以上。RIA试剂盒和CLIA试剂盒检测出正常人血清样本的特异度分别为98.11%（104/106）和99.06%（105/106）。癌症患者与正常人hVEGF测定值相比，差异均有统计学意义（t=−16.695~−14.920，均P<0.01）。 结论 2种试剂盒各项技术指标较好，其中CLIA试剂盒的分析灵敏度和特异度更高，在早期肺癌和结直肠癌的诊断中具有一定的临床价值。

Development of two kinds of human vascular endothelial growth factor immunoassay kits and their diagnostic value in early tumor

Objective To prepare human vascular endothelial growth factor (hVEGF) radioimmunoassay (RIA) and chemiluminescence immunoassay (CLIA) kits and evaluate their clinical value in diagnosis of early lung cancer and colorectal cancer. Methods A detection method was established according to the technical indices of the kits. The techniques for coating magnetic particles and 125iodine and acridine ester labeling with antibody were evaluated. Sensitivity, precision, and recovery of the kits were determined. The sensitivity and specificity of the kits in the diagnosis of early cancers (lung and colorectal cancers) were evaluated by testing cancer and normal serum samples. Independent sample t-test was used for inter-group comparison. Results The optimal ratios were as follows: 1 mL of magnetic particles to 1 mg of the antibody, 55.5 MBq 125iodine to 0.1 mg of the antibody, and 25 μg acridine ester to 0.2 mg of the antibody. The detection sensitivities of RIA and CLIA kits were 17.6 and 9.2 pg/mL respectively. For precision, the CLIA kit had slightly higher intra-batch variability and lower inter-batch variability than the RIA kit. The mean recovery levels of the RIA and CLIA kits were 103.28% and 101.85% respectively, and the latter had higher detection accuracy. The clinical sensitivity and specificity in the diagnosis of lung cancer and colorectal cancer were all above 90%. RIA kit and CLIA kit showed that the specificity of hVEGF in normal serum samples was 98.11% (104/106) and 99.06% (105/106), respectively.. There were significant differences in hVEGF between cancer patients and normal subjects (t=−16.695–−14.920, all P<0.01). Conclusions The technical indices of the hVEGF RIA and CLIA kits were good. The CLIA kit had higher sensitivity and specificity than the RIA kit and has clinical value in screening and auxiliary diagnosis of early lung and colorectal cancers.