竞争酶联免疫吸附试验检测人血管抑素的方法建立及其应用研究

Establishment and clinical application of the ELISA for plasma angiostatin examination

目的 研究建立竞争酶联免疫吸附试验 (竞争ELISA)检测血管抑素的方法 ,并探讨其临床应用价值。方法 以重组人血管抑素作为抗原免疫家兔 ,制备抗人血管抑素多克隆抗体 ,建立竞争ELISA法 ,并检测 6 1例肿瘤患者和 2 0名正常献血员血浆中血管抑素含量。结果 竞争ELISA法的灵敏度为 5mg/L ,批内和批间变异分别为 2 7%～ 3 2 %和 4 9%～ 5 1% ,平均回收率为10 0 4 %。血浆血管抑素含量在 2 0名献血员中为 2 7 2 2± 7 4 4mg/L ,2 8例胃癌患者为 5 7 15±2 9 90mg/L ,6例乳腺癌为 6 7 5 1± 2 0 88mg/L ,6例肝癌为 92 0 3± 18 72mg/L ,11例急性淋巴细胞性白血病为 4 1 72± 9 99mg/L ,10例其他肿瘤 (2例胆囊癌 ,3例结肠癌 ,2例前列腺癌 ,3例卵巢癌 )为 78 78± 2 0 4 4mg/L ,均显著高于正常人 (P <0 0 5或 0 0 1)。结论 建立的竞争ELISA为临床提供了一种简单、灵敏和特异地检测人血管抑素的方法 ,并可作为肿瘤诊断的一种新检测指标。

Objective To establish a ELISA to quantify human plasma angiostatin Methods With the rh angiostatin we immunized rabbit and acquired polycolonal antibodies against human angiostatin With these Antibodies, we established the competitive ELISA that could be used to quantify the human plasma angiostatin Results The assay sensitivity was 5 mg/L, coefficient of variation for inner batch and inter batch were 2 7% ～ 3 2% and 4 9% ～ 5 1% respectively The recovery was 100 4% The plasma level of An in normal control was 27 22 ±7 44 mg/L The plasma level of An was 57 15±29 90 mg/L in 28 patients of stomach cancer, 67 51±20 88 mg/L in 6 patients of breast cancer, 92 03±18 72 mg/L in 6 patients of liver cancer, 41 72±9 99 mg/L in 11 patients acute leukemia,and 78 28±20 44 mg/L in others (2 cases of gallbladder carcinoma, 3 patients of colon cancer, 2 patients of prostate carcinoma and 3 patients of ovary cancer) Conclusions Competitive ELISA for plasma angiostatin examination is a simple assay that can evaluate plasma angiostatin level accurately and provided a new method for diagnosis of cancer patients