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用于检测和定量选定致龋菌和牙周细菌的快速多重实时聚合酶链反应方法

Rapid Multiplex Real-Time PCR Method for the Detection and Quantification of Selected Cariogenic and Periodontal Bacteria.

作者信息

Lochman Jan, Zapletalova Martina, Poskerova Hana, Izakovicova Holla Lydie, Borilova Linhartova Petra

机构信息

Department of Biochemistry, Faculty of Science, Masaryk University, Kotlarska 2, 611 37 Brno, Czech Republic.

Department of Pathophysiology, Faculty of Medicine, Masaryk University, Kamenice 5, 625 00 Brno, Czech Republic.

出版信息

Diagnostics (Basel). 2019 Dec 22;10(1):8. doi: 10.3390/diagnostics10010008.

Abstract

Dental caries and periodontal diseases are associated with a shift from symbiotic microbiota to dysbiosis. The aim of our study was to develop a rapid, sensitive, and economical method for the identification and quantification of selected cariogenic and periodontal oral bacteria. Original protocols were designed for three real-time multiplex PCR assays to detect and quantify the ratio of 10 bacterial species associated with dental caries ("cariogenic" complex) or periodontal diseases (red complex, orange complex, and ). A total number of 60 samples from 30 children aged 2-6 years with severe early childhood caries and gingivitis were tested. In multiplex assays, the quantification of total bacterial (TB) content for cariogenic bacteria and red complex to eliminate differences in quantities caused by specimen collection was included. The mean counts for the TB load and that of ten evaluated specimens corresponded to previously published results. We found a significant difference between the microbial compositions obtained from the area of control and the affected teeth ( < 0.05). Based on this comprehensive microbiological examination, the risk of dental caries or periodontal inflammation may be determined. The test could also be used as a tool for behavioral intervention and thus prevention of the above-mentioned diseases.

摘要

龋齿和牙周疾病与共生微生物群向生态失调的转变有关。我们研究的目的是开发一种快速、灵敏且经济的方法,用于鉴定和定量选定的致龋性和牙周口腔细菌。设计了原始方案用于三种实时多重聚合酶链反应(PCR)检测,以检测和定量与龋齿相关的10种细菌物种(“致龋性”复合体)或牙周疾病(红色复合体、橙色复合体等)的比例。对30名2至6岁患有严重幼儿龋齿和牙龈炎的儿童的60个样本进行了检测。在多重检测中,包括对致龋细菌和红色复合体的总细菌(TB)含量进行定量,以消除样本采集导致的数量差异。TB负荷和十个评估样本的平均计数与先前发表的结果相符。我们发现,从对照区域和患牙获得的微生物组成之间存在显著差异(P<0.05)。基于这种全面的微生物学检查,可以确定龋齿或牙周炎症的风险。该检测还可作为行为干预的工具,从而预防上述疾病。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/427c/7168300/65e93511cc11/diagnostics-10-00008-g001.jpg

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