CAJ | 학술논문

目的利用变性梯度凝胶电泳(DGGE)技术观察牙周牙髓联合病变患牙牙周组织和根管中原始菌群分布状况及其异同,并通过克隆测序技术来试图探讨两部位可能存在的优势菌.方法从13例牙周牙髓联合病变患牙分别采集患牙根尖1/3处牙周细菌和根管牙髓细菌,提取细菌总DNA,扩增16S rRNA基因可变区,再进行变性梯度凝胶电泳.应用SPSS17.0软件和Quantity One软件对DGGE图谱菌种条带进行统计学分析和聚类分析.对DGGE凝胶中有代表性的条带进行回收和克隆测序.结果两取菌部位的菌种条带数间有明显的统计学差异(P＜0.01),但二者之间无正相关性.二者间的相似系数为13.1％～62.5％.牙周牙髓联合病变患牙根尖区1/3处牙周菌属可能有弯曲菌属(Campylobacter)、梭杆菌属(Fusobacterium)、奈瑟菌属(Neisseria)等,该处对应根管中菌属可能有优杆菌属(Mogibacterium)、棒状杆菌属(Corynebacterium)、放线菌属(Actinomyces)等.结论牙周牙髓联合病变(牙周来源)中牙周组织和邻近根管牙髓组织中菌种在数目和结构上有明显不同.该病变牙周组织和根管中可能存在目前尚未被认知的优势菌种.
목적 이용변성제도응효전영(DGGE)기술관찰아주아수연합병변환아아주조직화근관중원시균군분포상황급기이동,병통과극륭측서기술래시도탐토량부위가능존재적우세균.방법 종13례아주아수연합병변환아분별채집환아근첨1/3처아주세균화근관아수세균,제취세균총DNA,확증16S rRNA기인가변구,재진행변성제도응효전영.응용SPSS17.0연건화Quantity One연건대DGGE도보균충조대진행통계학분석화취류분석.대DGGE응효중유대표성적조대진행회수화극륭측서.결과 량취균부위적균충조대수간유명현적통계학차이(P＜0.01),단이자지간무정상관성.이자간적상사계수위13.1％～62.5％.아주아수연합병변환아근첨구1/3처아주균속가능유만곡균속(Campylobacter)、사간균속(Fusobacterium)、내슬균속(Neisseria)등,해처대응근관중균속가능유우간균속(Mogibacterium)、봉상간균속(Corynebacterium)、방선균속(Actinomyces)등.결론 아주아수연합병변(아주래원)중아주조직화린근근관아수조직중균충재수목화결구상유명현불동.해병변아주조직화근관중가능존재목전상미피인지적우세균충.
Objective To compare the bacterial community profiles present in periodontium and root canals of the same tooth diagnosed as combined periodontal-endodontic lesions by using denaturing gradient gel electrophoresis (DGGE).Methods Samples were collected from 13 extracted teeth with advanced periodontitis,endodontic samples from root tip 1/3 root canal,and periodontal samples from the corresponding neighboring periodontium.Genomic DNA was collected for the following universal bacterial primersPCR.The PCR products were then loaded on the DGGE gels to gain separate bands.The typical DGGE bands were excised,PCR-cloned and sequenced.Results The number of bands,which was indicative of the number of bacterial species,was compared intra-group (periodontal and pulpal specimen from the same tooth).The difference was statistically significant (P＜0.01),but there was no positive correlation between them.The similarity (Dice Coefficient) between them was 13.1％-62.5％.Taxa identified through BLAST (≥98％ identity) were Campylobacter,Fusobacterium,Neisseria,et al in the periodontium,and Mogibacterium,Corynebacterium,Actinomyces,et al in the root canals.Conclusion Common bacteria existed between them,but not all of the periodontal bacteria would appear in neighboring root canal; and the bacteria in the root canal are not completely from neighboring periodontal tissue.The original bacteria in the root canals may resuscitate and enrich the bacterial community.In combined periodontal-endodontic lesions (periodontal source),it is probable that new species existed to be confirmed either in the periodontium or in the root canal.