Robins S P, Woitge H, Hesley R, Ju J, Seyedin S, Seibel M J
Rowett Research Institute, Bucksburn, Aberdeen, Scotland.
J Bone Miner Res. 1994 Oct;9(10):1643-9. doi: 10.1002/jbmr.5650091019.
Several studies in recent years have shown that the pyridinium crosslinks of collagen provide good urinary markers of collagen degradation, primarily reflecting bone resorption. Most studies, however, were based on time-consuming HPLC assays of the crosslinks. We now describe the development of an immunoassay (ELISA) based on a monoclonal antibody for free deoxypyridinoline (Dpd) and its use in healthy individuals and patients with bone-related disorders to measure the urinary excretion of Dpd as an improved assessment of bone resorption rate. The Dpd antibody exhibited less than 1% cross-reaction with free pyridinoline and was shown to react only with free Dpd in urine, having no significant interaction with peptide forms of the crosslinks. The intra- and interassay variations were less than 10 and 15%, respectively. A total of 402 urine samples from patients and healthy volunteers were analyzed by both the immunoassay and HPLC. The ELISA results were highly correlated with those for total Dpd measured by HPLC over the full range of sample groups (r = 0.95). In normal adults, the excretion of Dpd (mean +/- SD) was 4.7 +/- 1.6 nmol/mmol creatinine, with about fivefold higher excretion rates in children. For 31 osteoporotic patients, the ELISA Dpd values (median 6.7; range 3.0-13.5 nmol/mmol Cr) were significantly higher (p < 0.0001) than the corresponding values for age- and sex-matched controls (median 4.0; range 1.8-7.4). The difference between the groups was similar for total Dpd by HPLC (osteoporotic: mean 12.8, range 4.8-30.7; controls: 6.6, range 3.0-18.1; p < 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS)
近年来的多项研究表明，胶原蛋白的吡啶交联物可作为反映胶原蛋白降解的良好尿液标志物，主要体现骨吸收情况。然而，大多数研究基于耗时的交联物高效液相色谱（HPLC）分析。我们现描述一种基于游离脱氧吡啶啉（Dpd）单克隆抗体的免疫测定法（酶联免疫吸附测定，ELISA）的开发，以及该方法在健康个体和患有骨相关疾病患者中的应用，以测量Dpd的尿排泄量，作为评估骨吸收速率的一种改进方法。Dpd抗体与游离吡啶啉的交叉反应小于1%，且仅与尿液中的游离Dpd发生反应，与交联物的肽形式无明显相互作用。批内和批间变异分别小于10%和15%。通过免疫测定法和HPLC对来自患者和健康志愿者的总共402份尿液样本进行了分析。在整个样本组范围内，ELISA结果与HPLC测定的总Dpd结果高度相关（r = 0.95）。正常成年人中，Dpd的排泄量（均值±标准差）为4.7±1.6 nmol/mmol肌酐，儿童的排泄率约高五倍。对于31名骨质疏松症患者，ELISA法测得的Dpd值（中位数6.7；范围3.0 - 13.5 nmol/mmol肌酐）显著高于年龄和性别匹配的对照组（中位数4.0；范围1.8 - 7.4）（p < 0.0001）。通过HPLC测定的总Dpd，两组之间的差异相似（骨质疏松症患者：均值12.8，范围4.8 - 30.7；对照组：6.6，范围3.0 - 18.1；p < 0.0001）。（摘要截短于250字）
