高致龋变形链球菌寡脱氧核苷酸探针的设计及有效性检测  

Identification of oligodeoxy nucleotide probe for high cariogenic Streptococcus mutans

作　　者：吴文[1] 耿海霞[1] 郭青玉[2] 

机构地区：[1]济宁医学院口腔系,山东济宁272013 [2]西安交通大学口腔医院口腔内科,陕西西安710004

出　　处：《临床口腔医学杂志》2010年第5期263-266,共4页Journal of Clinical Stomatology

摘　　要：目的:设计变形链球菌ATCC25175毒力因子gtfB寡脱氧核苷酸探针,并进行敏感性和特异性检测。方法:采用斑点杂交法,将体外合成的变形链球菌毒力因子gtfB寡脱氧核苷酸探针与经PCR扩增的S.M及7种口腔常见细菌DNA进行杂交检测,然后,使用该探针测试来源于髙龋和无龋个体的唾液样本。结果:探针检测同源性序列敏感性达到1ng水平;与7株口腔常见菌种的杂交结果不存在菌株间的交叉反应,具有较高的特异性。结论:变形链球菌毒力因子gtfB寡脱氧核苷酸探针敏感性高而且特异性好,化学性质稳定。同时,观察临床唾液样本测试结果发现,其阳性率与个体龋患现状基本一致。可以进一步考察该探针检测结果与临床病例龋患风险的一致性,为今后用于大规模流行病学调查中龋高危人群的筛查奠定基础。Objective：To design the digoxigenin-labelled 50 bp oligonucleotide probe,complementary to the virulence factor of Streptococcus mutans GS-5 gtfB,synthesize by AuGCT Biotechnology Company,and detect its sensitivity and specificity. Method：The PCR amplified DNA of Streptococcus mutans ATCC25175 and other conventional bacteria in hu- man mouth was dot-blotted with the oligonucleotide probe.Then test the saliva sample from the high decayed tooth individaul or the non-decayed tooth individual with the probe.observes thet result to try to discover the relations between the positive result and the situation of the decayed tooth in the individual. Result：The gtfB DNA probe of Streptococcus mutans could detect homologous target DNA fixed to nylon membrane at 1ng sensitivity. It could detect effectively Streptococcus mutans with no cross-reaction with six other oral bacteria. Conclusion：The sensitivity of Streptococcus mutans virulence factor gtfB oligonucleotide probe is not only high but also specific,and the chemical properties is stable.At the same time, using this probe test to the saliva sample which is gathered from the individual of different condition condition in caries.we can see that the probe test result is related with the present situation of the individual caries. So it is worth of studying the consistency between the probe test results and the caries risk from clinical cases. So that it can be used in large scale epi- demic investigation to screen caries susceptible people. At the same time this method makes a base of live examination for Streptococcus mutans.

关 键 词：变形链球菌 毒力因子gtfB 探针 

分 类 号：R780.2[医药卫生—口腔医学]