Assessment of a novel screening test for neutrophil collagenase activity in the diagnosis of periodontal diseases
Assessment of a novel screening test for neutrophil collagenase activity in the diagnosis of periodontal diseases
复制标题
评估牙周病诊断中性粒细胞胶原酶活性的新型筛查试验
DOI：
10.1902/jop.1999.70.11.1292
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发表时间：
1999-11-01
影响因子：
4.3
通讯作者：
McCulloch, CAG
中科院分区：
文献类型：
--
作者：
Mancini, S;Romanelli, R;McCulloch, CAG
背景：龈沟液（GCF）中活性中性粒细胞胶原酶（MMP-8）水平升高与进行性牙周炎有关。测量 GCF 中的这种酶有助于诊断。然而，目前具有足够灵敏度来检测全口 GCF 中胶原酶的检测使用放射性标记底物，并且需要几天才能完成。为了提供更快速的胶原酶活性分析，更好地适应临床研究，我们开发并验证了一种基于生物素化胶原消化产物的化学发光检测的新型测定（可溶性生物素化胶原测定：SBA）。方法：通过对来自 35 个中性粒细胞制剂和来自牙周炎患者的 41 个 GCF 样本的酶进行平行分析，评估新型 SBA 测定与放射性胶原底物测定的一致性，然后进行 Pearson 相关性分析分析。为了测试该测定是否适当地测量了 MMP-8 活性，在与特定胶原酶阻滞剂一起孵育后评估了酶活性。我们在 125 名成人牙周炎患者、5 名早发性牙周炎患者、1 名无牙颌患者和 32 名无牙周炎对照患者的横断面和纵向分析中检查了 SBA 的诊断效用。结果：该检测检测到的重度牙周炎患者比稳定牙周炎患者高 18 倍，并下降到
Background: Increased levels of active neutrophil collagenase (MMP-8) in the gingival crevicular fluid (GCF) are associated with progressive periodontitis. The measurement of this enzyme in GCF could facilitate diagnosis. However, assays with sufficient sensitivity to detect collagenase in whole-mouth GCF currently use radiolabeled substrates and require several days to complete. To provide more rapid analyses of collagenase activity that are better adapted to clinical studies, we developed and validated a novel assay (soluble biotinylated-collagen assay: SBA) based on chemiluminescent detection of biotinylated collagen digestion products.Methods: The concordance of the novel SBA assay with a radioactive collagen substrate assay was assessed by parallel analyses of enzyme from 35 neutrophil preparations and from 41 samples of GCF from periodontitis patients, followed by Pearson correlation analysis. To test whether the assay appropriately measured MMP-8 activity, enzyme activity was assessed after incubation with specific collagenase blockers. We examined the diagnostic utility of the SBA in cross-sectional and longitudinal analyses of 125 patients with adult periodontitis, 5 patients with early-onset: periodontitis, 1 edentulous patient, and in 32 control patients without periodontitis.Results: The assay detected 18-fold higher in severe periodontitis than in stable periodontitis and decreased to