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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 gtfB oligodeoxynucleotide probe of Streptococcus mutans ATCC25175 and to detect its sensitivity and specificity. Methods: Speckled hybridization was used to detect the S. mutans virulence factor gtfB oligodeoxynucleotide probe hybridized in vitro with SM and seven kinds of common bacterial DNA amplified by PCR. Then, Saliva samples from caries and caries-free individuals. Results: The sensitivity of the probe to detect homology was 1ng. The results of hybridization with 7 strains of common oral bacteria showed no cross-reaction between isolates and had high specificity. Conclusion: Streptococcus mutans virulence factor gtfB oligodeoxynucleotide probe is highly sensitive, specific and chemically stable. At the same time, observe the clinical saliva sample test results found that the positive rate and the status of individual caries basically the same. The consistency between the probe test results and the caries risk in clinical cases can be further investigated, which lays the foundation for the future screening of high-risk caries in large-scale epidemiological survey.