Development of a diagnostic tool for periodontal disease by utilizing the monoclonal antibody directed the specific protease from periodontopathogensis bacteria

制备了抗牙龈卟啉单胞菌胰酶样酶(Pase-C)的单抗(mAb-PC)。中和实验表明，单抗-PC与Pase-C结合，使Pase-C远离酶的活性部位，并抑制其酶活性。牙龈假单胞菌的其他类胰酶(PASE-B和PASE-S)不与该抗体反应。通过酶联免疫吸附试验和斑点印迹分析，mAbPC可识别所有牙周病原菌，但不识别其他可能的牙周病致病微生物，包括福赛氏杆菌和齿密螺旋体。经统计学分析，Pase-C水平与牙龈假单胞菌水平无相关性。但对探查时未发现出血的“BOP阴性组”进行分析时，这些水平显著相关。“BOP阳性组”Pase-C水平高于“BOP阴性组”，而Pase-C水平两组间差异无统计学意义。由于出血倾向在一定程度上反映了牙周炎局部的炎症活动，故牙周液标本中Pase-C的检测有可能用于成人牙周炎的诊断和预后判断。

A monoclonal antibody (mAb-PC) was produced against a trypsin-like enzyme (Pase-C) from Porphyromonas gingivalis. By the neutrolizing experiment, it was revealed that the mAb-PC bound Pase-C apart from the acive site of the enzyme and ihibited its enzyme activity. Other trypsin-like enzymes (Pase-B and Pase-S) from P.gingivalis did not react with this antibody. By ELISA or dot blotting analysis, mAb-PC recognized all strains of P.gingivalis tested, but did not recognized other putative periodontopathogenic organisms including Bacteroides forsythus and Treponema denticola.Fifty samples of gingival crevicular fluid (GCF) were collected from individuals with adult periodontitis, and the levels of Pase-C and P.gingivalis antigens in GCF were measured by a quantitative ELISA,using mAb-PC and anti-P.gingivalis polyclonal antibody, respectively. By the stastistic analysis, it was found that the Pase-C level was not correlated with the P.gingivalis level. However, when "BOP negative group" which bleeding was not found on probing, was subjected to the analysis, these levels were correlated singnificantly. The Pase-C level of "BOP positive group" was higher than that of "BOP negative group" , whereas on the P.gingivalis level, there was not any difference between these groups. Since the bleeding tendency is considered to reflect the inflammatory activity in the periodontal site to some extent, it was suggested that the detection of Pase-C in GCF sample had the potential to be used for the diagnosis and the prediction of prognosis in adult periodontitis.