Infection frequency of Epstein-Barr virus in subgingival samples from patients with different periodontal status and its correlation with severity of periodontal lesion

Abstract

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. 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 amplicons were further identified by RFLP with endonucleases 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. 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 endonuclease 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. Subgingival infection with either EBV-1 or EBV-2 is closely associated with chronic periodontitis. Infection of EBV may not correlate directly with severity of periodontitis.