Molecular and clinical analyses of Helicobacter pylori colonization in inflamed dental pulp
Molecular and clinical analyses of Helicobacter pylori colonization in inflamed dental pulp
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DOI：
10.1186/s12903-018-0526-2
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发表时间：
2018-04-16
期刊：
影响因子：
2.9
通讯作者：
Nakano, Kazuhiko
中科院分区：
文献类型：
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作者：
Nomura, Ryota;Ogaya, Yuko;Nakano, Kazuhiko
背景：近年来，牙髓被认为是儿童幽门螺杆菌(H.Pylori)感染的一个可能来源。我们以前开发了一种新的检测幽门螺杆菌的聚合酶链式反应系统，使用基于48株幽门螺杆菌的全基因组信息构建的引物集，具有高特异性和敏感性。以系列稀释度的幽门螺杆菌基因组DNA为模板，该体系具有很高的灵敏度，检测下限为1-10个细胞。然而，从炎症牙髓标本中检测幽门螺杆菌细菌DNA的检测下限较低(10(2)-10(3)个细胞)。因此，我们进一步改进了套式聚合酶链式反应方法，该方法比以前的单一聚合酶链式反应方法更灵敏。此外，我们还检测了幽门螺杆菌在炎症牙髓组织中的分布和毒力。方法：根据48株幽门螺杆菌的全基因组信息设计引物，构建套式聚合酶链式反应体系。利用从炎症牙髓标本中分离的幽门螺杆菌基因组DNA和细菌DNA，该套式聚合酶链式反应系统的检测下限为1-10个细胞。接下来，用套式聚合酶链式反应系统检测131例炎症牙髓标本中幽门螺杆菌的分布。此外，还研究了幽门螺杆菌的检测与根管感染牙齿的临床信息之间的关系。结果：38.9%的炎症牙髓标本在套式聚合酶链式反应系统中检出H.幽门螺杆菌主要在乳牙中检出，而不是恒牙。此外，每隔一周或两周对同一颗牙齿进行两次发炎牙髓采样，结果显示在两次采样中的大多数样本中都检测到幽门螺杆菌。此外，幽门螺杆菌菌株对人牙本质成纤维细胞具有粘附性。结论：我们的结果表明，大约40%的病例中幽门螺杆菌通过与人牙本质成纤维细胞的粘附来定植炎症牙髓。
Background: Recently, dental pulp has been considered a possible source of infection of Helicobacter pylori (H. pylori) in children. We previously developed a novel PCR system for H. pylori detection with high specificity and sensitivity using primer sets constructed based on the complete genome information for 48 H. pylori strains. This PCR system showed high sensitivity with a detection limit of 1-10 cells when serial dilutions of H. pylori genomic DNA were used as templates. However, the detection limit was lower (10(2)-10(3) cells) when H. pylori bacterial DNA was detected from inflamed pulp specimens. Thus, we further refined the system using a nested PCR method, which was much more sensitive than the previous single PCR method. In addition, we examined the distribution and virulence of H. pylori in inflamed pulp tissue.Methods: Nested PCR system was constructed using primer sets designed from the complete genome information of 48 H. pylori strains. The detection limit of the nested PCR system was 1-10 cells using both H. pylori genomic DNA and bacterial DNA isolated from inflamed pulp specimens. Next, distribution of H. pylori was examined using 131 inflamed pulp specimens with the nested PCR system. In addition, association between the detection of H. pylori and clinical information regarding endodontic-infected teeth were investigated. Furthermore, adhesion property of H. pylori strains to human dental fibroblast cells was examined.Results: H. pylori was present in 38.9% of inflamed pulp specimens using the nested PCR system. H. pylori was shown to be predominantly detected in primary teeth rather than permanent teeth. In addition, samplings of the inflamed pulp were performed twice from the same teeth at 1- or 2-week intervals, which revealed that H. pylori was detected in most specimens in both samplings. Furthermore, H. pylori strains showed adhesion property to human dental fibroblast cells.Conclusion: Our results suggest that H. pylori colonizes inflamed pulp in approximately 40% of all cases through adhesion to human dental fibroblast cells.