Department of Restorative Dentistry, Division of Endodontics, Piracicaba Dental School, State University of Campinas, Av Limeira 901, Bairro Areiao, Piracicaba, São Paulo, Brazil.
Clin Oral Investig. 2021 Sep;25(9):5171-5179. doi: 10.1007/s00784-021-03823-w. Epub 2021 Feb 9.
To evaluate strains of Enterococcus faecalis isolated from endodontic failures cases for (a) presence of virulence genes, namely, gelatinase production (gelE), surface protein (esp), collagen-binding adhesin (ace), cytolysin activator (cylA), E. faecalis antigen A (efaA) and aggregation substance (asa), all by using PCR; (b) biofilm formation capacity; and (c) activity of gelatinase and β-lactamase.
Twenty-five strains of E. faecalis were tested. The DNA extracted from these strains was used for identification of virulence genes by PCR and 1% agarose gel. Biofilm formation was performed on polystyrene microplates by using the violet crystal staining method. For assessment of the gelatinase activity, inoculum of pure cultures was deposited in tubes containing gelatin and a nutrient broth, whereas nitrocefin disks were used to assess the β-lactamase action.
The virulence genes efaA and cylA were detected in 100% of the strains, whereas gelE was present in 84%, ace in 68%, esp in 56% and asa in 48%. Four strains had no biofilm formation, 17 had poor formation and four had moderate formation. Gelatinase production was observed in three strains and β-lactamase resistance in five strains of E. faecalis.
Diverse patterns of virulence gene detection were observed among the E. faecalis strains, with predominance of those capable of forming biofilm. A few strains have been found to hydrolyze gelatin proteins, whereas β-lactamase resistance was detected in different isolates.
To understand the influence of virulence factors in E. faecalis on the host heath status.
评估从根管治疗失败病例中分离出的粪肠球菌菌株(a)是否存在毒力基因,即明胶酶产生(gelE)、表面蛋白(esp)、胶原结合黏附素(ace)、细胞毒素激活剂(cylA)、粪肠球菌抗原 A(efaA)和聚集物质(asa),所有这些都通过 PCR 进行检测;(b)生物膜形成能力;和(c)明胶酶和β-内酰胺酶的活性。
测试了 25 株粪肠球菌。从这些菌株中提取的 DNA 用于通过 PCR 和 1%琼脂糖凝胶鉴定毒力基因。通过使用紫晶染色法在聚苯乙烯微孔板上进行生物膜形成。为了评估明胶酶活性,将纯培养物的接种物沉积在含有明胶和营养肉汤的管中,而使用硝噻吩二乙酸酯(nitrocefin)盘来评估β-内酰胺酶的作用。
100%的菌株检测到毒力基因 efaA 和 cylA,84%的菌株检测到 gelE,68%的菌株检测到 ace,56%的菌株检测到 esp,48%的菌株检测到 asa。有 4 株菌没有形成生物膜,17 株菌形成的生物膜较差,4 株菌形成的生物膜适中。观察到 3 株粪肠球菌产生明胶酶,5 株粪肠球菌产生β-内酰胺酶耐药性。
在粪肠球菌菌株中观察到不同的毒力基因检测模式,以能够形成生物膜的菌株为主。发现少数菌株能够水解明胶蛋白,而不同分离株中检测到β-内酰胺酶耐药性。
了解粪肠球菌中毒力因子对宿主健康状况的影响。