Arakawa S, Hata S, Ishikawa I, Tsuchida N
Department of Periodontology, Faculty of Dentistry, Tokyo Medical and Dental University, Japan.
Arch Oral Biol. 1990;35 Suppl:93S-96S. doi: 10.1016/0003-9969(90)90136-x.
Actinobacillus actinomycetemcomitans has been implicated in the aetiology of juvenile periodontitis and advanced destructive periodontitis. Levels of IgG antibody against A. actinomycetemcomitans in peripheral blood sera of patients with advanced destructive periodontitis are high, as are those against Bacteroides gingivalis. To clone the genes of antigens reactive with sera of such patients, a library of the A. actinomycetemcomitans strain Y4 DNA in lambda L47 was constructed and then screened, using an immunochemical detection method, with serum from a patient with the advanced disease. Six clones from among nearly 1000 reacted with the serum and also with that of another patient. They were designated 3, 4, 6, 7, 8 and 9. Restriction enzyme and Southern blot analyses indicated that clones 8 and 9 were identical and that all the clones were overlapping because they shared in common the 4 and 5 kbp HincII DNA fragments of A. actinomycetemcomitans. The cloned DNA fragment hybridized to the DNA of two other strains of A. actinomycetemcomitans but not to those of six periodontopathic bacteria examined. These findings suggest that a DNA sequence encoding an A. actinomycetemcomitans strain Y4 antigen strongly reactive with sera of patients with advanced destructive periodontitis was cloned. This sequence is present specifically in A. actinomycetemcomitans but not in other bacteria isolated from patients with periodontal diseases. Thus, the cloned DNA could serve as a probe for the diagnosis of periodontitis.
伴放线放线杆菌与青少年牙周炎及重度破坏性牙周炎的病因学有关。重度破坏性牙周炎患者外周血血清中抗伴放线放线杆菌的IgG抗体水平较高，抗牙龈拟杆菌的抗体水平也较高。为了克隆与这类患者血清反应的抗原基因，构建了伴放线放线杆菌Y4菌株DNA在λL47中的文库，然后用免疫化学检测方法，用一名重度患者的血清进行筛选。近1000个克隆中有6个与该血清以及另一名患者的血清发生反应。它们被命名为3、4、6、7、8和9。限制性内切酶和Southern印迹分析表明，克隆8和9相同，所有克隆都相互重叠，因为它们共享伴放线放线杆菌4和5kbp的HincII DNA片段。克隆的DNA片段与另外两株伴放线放线杆菌的DNA杂交，但与检测的6种牙周病原菌的DNA不杂交。这些发现表明，克隆到了一个编码与重度破坏性牙周炎患者血清强烈反应的伴放线放线杆菌Y4菌株抗原的DNA序列。该序列特异性地存在于伴放线放线杆菌中，而不存在于从牙周病患者分离出的其他细菌中。因此，克隆的DNA可作为牙周炎诊断的探针。
