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设计和验证一种 DNA 微阵列,用于分析不同口腔样本和牙种植体中的细菌群落的系统发育。

Design and validation of a DNA-microarray for phylogenetic analysis of bacterial communities in different oral samples and dental implants.

机构信息

Department of Pharmacy and Biotechnology, University of Bologna, Bologna, Italy.

Institute of Biomedical Technologies - National Research Council, Segrate, Milan, Italy.

出版信息

Sci Rep. 2017 Jul 24;7(1):6280. doi: 10.1038/s41598-017-06743-6.

DOI:10.1038/s41598-017-06743-6
PMID:28740183
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5524749/
Abstract

The quali-quantitative characterization of the oral microbiota is crucial for an exhaustive knowledge of the oral ecology and the modifications of the microbial composition that occur during periodontal pathologies. In this study, we designed and validated a new phylogenetic DNA-microarray (OralArray) to quickly and reliably characterize the most representative bacterial groups that colonize the oral cavity. The OralArray is based on the Ligation Detection Reaction technology associated to Universal Arrays (LDR-UA), and includes 22 probe sets targeted to bacteria belonging to the phyla Firmicutes, Proteobacteria, Actinobacteria, Bacteroidetes, Fusobacteria, and Spirochaete. The tool is characterized by high specificity, sensitivity and reproducibility. The OralArray was successfully tested and validated on different oral samples (saliva, lingual plaque, supragingival plaque, and healing cap) collected from 10 healthy subjects. For each specimen, a microbial signature was obtained, and our results established the presence of an oral microbial profile specific for each subject. Moreover, the tool was applied to evaluate the efficacy of a disinfectant treatment on the healing caps before their usage. The OralArray is, thus, suitable to study the microbiota associated with various oral sites and to monitor changes arising from therapeutic treatments.

摘要

口腔微生物群落的定性-定量特征对于全面了解口腔生态学以及牙周病过程中微生物组成的变化至关重要。在本研究中,我们设计并验证了一种新的基于聚合酶链式反应(PCR)技术的口腔微生物群落 DNA 芯片(OralArray),以快速、可靠地鉴定口腔中最具代表性的细菌群落。OralArray 基于连接酶检测反应(Ligation Detection Reaction)技术与通用探针阵列(Universal Arrays)相结合,包含 22 个探针组,针对厚壁菌门(Firmicutes)、变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、拟杆菌门(Bacteroidetes)、梭杆菌门(Fusobacteria)和螺旋体门(Spirochaete)的细菌。该工具具有高度的特异性、灵敏度和重现性。我们成功地在 10 名健康受试者的不同口腔样本(唾液、舌背菌斑、龈上菌斑和愈合帽)上对 OralArray 进行了测试和验证。对于每个样本,我们都获得了一个微生物特征谱,我们的结果确定了每个受试者的口腔微生物特征谱。此外,该工具还用于评估消毒剂处理对愈合帽在使用前的消毒效果。因此,OralArray 适合研究与各种口腔部位相关的微生物群落,并监测治疗过程中产生的变化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/849e/5524749/e5c7cb68c696/41598_2017_6743_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/849e/5524749/276b86dbd0ad/41598_2017_6743_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/849e/5524749/4eaa6c3dc389/41598_2017_6743_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/849e/5524749/a5c4e4fabd92/41598_2017_6743_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/849e/5524749/d704267104be/41598_2017_6743_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/849e/5524749/246e0980922f/41598_2017_6743_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/849e/5524749/e5c7cb68c696/41598_2017_6743_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/849e/5524749/276b86dbd0ad/41598_2017_6743_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/849e/5524749/4eaa6c3dc389/41598_2017_6743_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/849e/5524749/a5c4e4fabd92/41598_2017_6743_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/849e/5524749/d704267104be/41598_2017_6743_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/849e/5524749/246e0980922f/41598_2017_6743_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/849e/5524749/e5c7cb68c696/41598_2017_6743_Fig6_HTML.jpg

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