摘要
目的建立多重PCR方法检测正畸治疗患者口腔中伴放线放线杆菌(Aa)、福塞氏类杆菌(Bf)、具核梭杆菌(Fn)、牙龈卟啉单胞菌(Pg)四种细菌,并与牙龈指数进行了相关性分析。方法选择55例正畸治疗至少2个月的患者为实验组,34例未带矫治器的牙周健康者为对照组,记录牙龈炎症指数,分别采集牙周袋最深处或龈沟液标本进行细菌DNA提取及聚合酶链式反应,同时以细菌的厌氧培养和生化反应鉴定为标准进行对比验证,并做了PCR的敏感性和特异性实验。结果建立的多重PCR最低可检测出1pg的细菌DNA,约相当于20个Aa、Fn和Pg,80个Bf;能扩增出Aa、Bf、Fn、Pg四种参考菌株的目的条带,而对EC的扩增没有目的条带;多重PCR法与常规细菌培养法对细菌的检测阳性率较为一致(P>0.05);多重PCR检测发现Aa、Fn、Pg在两组间的差异有统计学意义(P<0.05),对Bf的检测无统计学意义(P>0.05);Spearman等级相关分析PCR检出Aa、Fn、Pg的阳性率与牙龈指数之间存在明显的正相关关系(P<0.01);而Bf与牙龈指数之间未有明显的相关关系(P>0.05)。结论所建立的多重PCR有较高的敏感性和特异性,可同时检测龈沟液中四种牙周致病菌,观察因固定矫治器的使用而引起的牙周细菌的变化。
Objective To establish a multiplex PCR for detection of four species of bacteria in oral specimens from orthodontic patients, and analyse the correlation between presence of bacteria and ginival index. Methods Periodomal pocket specimens from fifty-five young patients who wore fixed orthodontic appliances and thirty-four healthy individuals who did not wear orthodontic appliances were collected and the bacteria DNAs were extracted, then polymerase chain reactions were used to amplify the target genes.Meanwhile, anaerobic culture and biochemical event were used to verify PCR results. The sensitivity and specificity of PCR were evaluated with the reference bacteria. In addition, the gingival indexes of each individual were recorded. Results The multiplex PCR was able to detect 1 pg bacteria DNAs corresponding to 20 cells of Aa, Fn and Pg as well as 80 cells of Bf. With the good specificity, four species of bacteria were produced target fragments and the E. coh was negative. The positive rate of PCR assay was coincident with the bacteria culture （ P 〉 0.05）. The differences between two groups were found to have statistical significance for detection of Aa, Fn and Pg（ P 〈 0.05）and have no significance for detecting Bf. The analysis of Spearman rank correlation indicated that the presences of Aa, Fn, Pg have the positive correlation with gingival indexes （ P 〈 0.01 ） and no correlation with Bf （ P 〉 0.05）. Conclusion With high sensitivity and specificity,the multiplex PCR can be used to detect Aa, Bf, Fn and Pg simultaneously and observe the bacteria changes induced by wearing fixed orthodontic appliances.
出处
《中国实验诊断学》
北大核心
2009年第11期1579-1582,共4页
Chinese Journal of Laboratory Diagnosis
基金
济南市科技局资助科研项目[济科合字2006-061040]
济南市卫生局资助项目[济卫科外发2004-36]
关键词
固定矫治器
多重PCR
伴放线放线杆菌
福塞氏类杆菌
具核梭杆菌
牙龈卟啉单胞菌
Fixed orthodontic appliances
Multiplex PCR
Actinobaeillns actinomycetemcomitans
Bacteroides forsythns
Fusobacterium nucleatum
Porphyromonas gingivalis
作者简介
作者简介：刘晓华，男，44岁，主要从事口腔医学的临床科研、教学工作。
通讯作者
