Molecular Identification of Candida albicans in Endodontic Retreatment Cases by SYBR Green I Real-time Polymerase Chain Reaction and its Association with Endodontic Symptoms.

Ahmad Nouroloyouni, Negar Moghaddam, Sarah Nuroloyuni, Amin Salem Milani, Hamid Reza Yavari, Ali Reza Majidi
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Abstract

Statement of the problem: Recent microbiological studies have expressed ever-increasing concerns about Candida albicans as a causal factor in the failure of endodontic treatments. Real-time quantitative polymerase chain reaction (qPCR), including the SYBR Green I system, is a technique in which a fluorescent dye is incorporated into the double-stranded DNA that is produced during DNA polymerase activity.

Purpose: This study aimed to determine the relative prevalence of Candida albicans in root canals of retreatment cases and its association with endodontic symptoms.

Materials and method: In the present cross-sectional/analytical study fifty subjects were selected. Clinical features and radiographic status of the teeth were also evaluated. After access cavity preparation, the retrieved material and dentinal chips removed from the root canal were transferred into 1.5-mL microtubes, followed by storage at -20ºC until used for DNA extraction. A DNeasy Tissue Kit was used to extract DNA using the DNeasy protocol for animal tissues. Master Plus SYBR Green I (Jena Bioscience, Germany) was used in a Rotor-gene Real-time PCR System for real-time PCR. The relationship between the presence of Candida albicans and the clinical and radiographic features were analyzed using McNemar's test.

Results: There was a significant relationship between the radiographic findings in endodontically treated teeth and the presence of Candida albicans. However, there was no significant relationship between the presence of Candida albicans and any of the clinical symptoms.

Conclusion: In spite of the limitations of this study, we concluded that Candida albicans was associated with root canal infections in endodontic retreatment cases, but there was no relationship between root canal infections and the clinical symptoms.

利用 SYBR Green I 实时聚合酶链反应对牙髓再治疗病例中的白色念珠菌进行分子鉴定及其与牙髓症状的关系
问题陈述:最近的微生物学研究表明,白色念珠菌是牙髓治疗失败的一个致病因素,这引起了越来越多的关注。实时定量聚合酶链反应(qPCR),包括 SYBR Green I 系统,是一种在 DNA 聚合酶活动过程中产生的双链 DNA 中加入荧光染料的技术。目的:本研究旨在确定再治疗病例根管内白色念珠菌的相对流行率及其与牙髓症状的关联:本横断面/分析性研究选取了 50 名受试者。研究还评估了牙齿的临床特征和影像学状况。在进行牙洞准备后,将取回的材料和从根管中取出的牙本质碎屑转移到 1.5 毫升的微管中,然后储存在 -20ºC 温度下,直到用于提取 DNA。使用 DNeasy 组织试剂盒按照动物组织的 DNeasy 方案提取 DNA。在 Rotor-gene Real-time PCR 系统中使用 Master Plus SYBR Green I(德国耶拿生物科学公司)进行实时 PCR。用 McNemar 检验分析了白色念珠菌的存在与临床和放射学特征之间的关系:结果:牙髓治疗后牙齿的影像学检查结果与白色念珠菌的存在有明显关系。然而,白色念珠菌的存在与任何临床症状之间均无明显关系:尽管这项研究有其局限性,但我们得出结论,在牙髓再治疗病例中,白色念珠菌与根管感染有关,但根管感染与临床症状之间没有关系。
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