针对导致牙龈脓肿的抗生素耐药菌,全面鉴定有效的噬菌体。
Fully Characterized Effective Bacteriophages Specific against Antibiotic-Resistant , the Causative Agent of Dental Abscess.
机构信息
Botany and Microbiology Department, Faculty of Science, Benha University, Benha 13511, Egypt.
Botany and Microbiology Department, Faculty of Science, Al-Azhar University, Nasr, Cairo 11884, Egypt.
出版信息
Medicina (Kaunas). 2024 Mar 19;60(3):501. doi: 10.3390/medicina60030501.
: is a primary pathogen responsible for dental abscesses, which cause inflammation and pain when trapped between the crown and soft tissues of an erupted tooth. Therefore, this study aims to use specific phages as an alternative method instead of classical treatments based on antibiotics to destroy multidrug-resistant bacteria for treating dental issues. : In the current study, twenty-five bacterial isolates were obtained from infected dental specimens; only five had the ability to grow on bile esculin agar, and among these five, only two were described to be extensive multidrug-resistant isolates. : Two bacterial isolates, A.R.A.01 [ON797462.1] and A.R.A.02, were identified biochemically and through 16S rDNA, which were used as hosts for isolating specific phages. Two isolated phages were characterized through TEM imaging, which indicated that _phage-01 had a long and flexible tail, belonging to the family , while . _phage-02 had a contractile tail, belonging to the family Genetically, two phages were identified through the PCR amplification and sequencing of the RNA ligase of phage vB_EfaS_HEf13, through which our phages shared 97.2% similarity with phage vB-EfaS-HEf13 based on BLAST analysis. Furthermore, through in silico analysis and annotations of the two phages' genomes, it was determined that a total of 69 open reading frames (ORFs) were found to be involved in various functions related to integration excision, replication recombination, repair, stability, and defense. In phage optimization, the two isolated phages exhibited a high specific host range with among six different bacterial hosts, where _phage-01 had a latent period of 30 min with 115.76 PFU/mL, while . _phage-02 had a latent period of 25 min with 80.6 PFU/mL. They were also characterized with stability at wide ranges of pH (4-11) and temperature (10-60 °C), with a low cytotoxic effect on the oral epithelial cell line at different concentrations (1000-31.25 PFU/mL). : The findings highlight the promise of phage therapy in dental medicine, offering a novel approach to combating antibiotic resistance and enhancing patient outcomes. Further research and clinical trials will be essential to fully understand the therapeutic potential and safety profile of these bacteriophages in human populations.
是一种主要的病原体,可导致牙脓肿,当被困在出牙的牙齿冠和软组织之间时,会引起炎症和疼痛。因此,本研究旨在使用特定噬菌体作为替代方法,而不是基于抗生素的经典治疗方法来破坏多药耐药菌,以治疗牙科问题。在目前的研究中,从感染的牙科标本中获得了 25 个细菌分离物;只有 5 个能够在胆汁七叶苷琼脂上生长,其中只有 2 个被描述为广泛的多药耐药分离物。两个细菌分离物,A.R.A.01 [ON797462.1]和 A.R.A.02,通过 16S rDNA 进行生化和鉴定,被用作分离特定噬菌体的宿主。通过 TEM 成像对两种分离的噬菌体进行了表征,表明噬菌体-01 具有长而灵活的尾巴,属于科,而噬菌体-02 具有收缩的尾巴,属于科。通过聚合酶链反应 (PCR) 扩增和测序噬菌体 vB_EfaS_HEf13 的 RNA 连接酶,对两种噬菌体进行了基因鉴定,通过 BLAST 分析,我们的噬菌体与噬菌体 vB-EfaS-HEf13 的相似度为 97.2%。此外,通过对两种噬菌体基因组的计算机分析和注释,确定总共发现 69 个开放阅读框 (ORF) 参与了与整合切除、复制重组、修复、稳定性和防御相关的各种功能。在噬菌体优化中,两种分离的噬菌体表现出与六种不同细菌宿主之间的高特异性宿主范围,其中噬菌体-01 的潜伏期为 30 分钟,效价为 115.76 PFU/mL,而噬菌体-02 的潜伏期为 25 分钟,效价为 80.6 PFU/mL。它们还具有在宽 pH(4-11)和温度(10-60°C)范围内的稳定性,在不同浓度(1000-31.25 PFU/mL)下对口腔上皮细胞系的细胞毒性作用较低。研究结果强调了噬菌体治疗在牙科医学中的应用前景,为对抗抗生素耐药性和改善患者预后提供了一种新方法。进一步的研究和临床试验对于充分了解这些噬菌体在人群中的治疗潜力和安全性至关重要。