摘要
目的了解慢性牙周炎(CP)龈下标本中不同基因型EB病毒(EBV)的感染率及其与CP病变程度的关系。方法采用EBV1和EBV2特异引物的套式聚合酶链反应(nPCR),分别检测65例CP患者、65例牙龈炎患者和24例牙周健康者龈下标本中的EBV1和EBV2。采用内切酶AfaⅠ和StuⅠ的限制性片段长度多态性分析(RFLP)进一步鉴定扩增产物。以牙周附着丧失和牙龈指数为观察指标,分析EBV感染与否及EBV1或EBV2感染与牙周炎严重程度的关系。结果所建立的nPCR可检测出0.01ng的EBV1DNA,所有标本两次nPCR检测结果均相同。497bp的EBV1扩增产物用AfaⅠ酶切后均可分为355bp和142bp两个片段,165bp的EBV2扩增产物用StuⅠ酶切后均可分为118bp和47bp两个片段。在CP、牙龈炎和牙周健康者标本中,EBV1阳性率分别为28.5%(74/260)、16.9%(44/260)和14.6%(14/96),EBV2分别为8.1%(21/260)、3.1%(8/260)和0,无一例标本中同时检出EBV1和EBV2,EBV总阳性率分别为36.5%(95/260)、20.0%(52/260)和14.6%(14/96)。EBV1、EBV2阳性率和EBV总阳性率在CP标本中均高于牙龈炎标本(P<0.05)和正常龈沟标本(P<0.01),但在牙龈炎与正常龈沟标本间差异无统计学意义(P>0.05)。未发现牙周炎患者EBV感染与否及EBV1或EBV2感染与附着丧失和牙龈指数有何关联。结论EBV1和EBV2感染均与CP密切相关。EBV感染与牙周炎严重程度无直接关系。
Objective To detect the infection frequencies of different Epstein-Barr virus （ EBV ） genotypes in subgingival samples of chronic periodontitis, and the correlation among infection with different genotypes and the severity of periodontal lesion. Methods Nested PCR （nPCR） with EBV-1 or EBV-2 specific primers was used to detect EBV-1 and EBV-2 in the subgingival samples from 65 chronic periodontitis patients, 65 gingivitis patients and 24 periodontal healthy individuals. The amplieons were further identified by RFLP with endonueleases Afa I and Stu I. By using periodontal attachment loss （AL） and gingival index （GI） as the observing in, the correlation of infection with different EBV genotypes and the severity of periodontal lesion were analyzed. Results 0.01 ng of EBV-1 DNA could be detectable by the established nPCR. All the samples showed the same detection results by two separated nPCR. All the EBV-1 amplification products （497 bp） by using endonuclease Afa I digestion could be divided into two fragments with 355 bp and 142 bp respectively. After endonuelease Stu I digestion, all the EBV-2 amplification products （ 165 bp） displayed two fragments with 118 bp and 47 bp respectively. In the samples of chronic periodontitis patients, gingivitis patients, and healthy periodontal tissues, the positive rates were 28.5% （74/260） ,16.9% （44/260）, and 14.6% （14/96） for EBV-1 ; and were 8.1% （21/260）, 3.1% （8/260）, and 0% for EBV-2 respectively, and the total EBV positive rates were 36. 5% （95/260）, 20.0% （52/260） and 14.6% （14/96） respectively. None of the positive samples was detectable for both the EBV-1 and EBV-2. The positive rates of EBV-1, EBV-2 and the total EBV positive rates in the chronic periodontitis samples were all higher than those in the gingivitis samples （all P 〈 0. 05 ） and healthy periodontal tissue samples （all P 〈 0.01 ） , without a significant difference between the gingivitis samples and healthy periodontal tissue samples （P 〉 0.05）. Infection of EBV or EBV-1 or EBV-2 in CP patients could not be associated with AL or GI. Conclusion Subgingival infection with either EBV-1 or EBV-2 is closely associated with chronic penodontitis. Infection of EBV may not correlate directly with severity of periodontitis.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2005年第45期3216-3220,共5页
National Medical Journal of China
基金
浙江省科技计划一般项目基金资助(N021107286)
