Detection of Streptococcus mutans and Streptococcus sobrinus in Human Dental Plaque Samples by Using Semi-Quantitative Real-Time Polymerase Chain Reaction

Puja Banduji Paunfase, Samynathan Ramkumar, Marappan Ganesan, Veeraraghavan Usha
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Abstract

Dental caries, a highly prevalent infectious disease in humans is caused by the bacterial plaque that coats the teeth surface and is a serious public health concern. Recently, the formation of dental plaque has been associated with the occurrence of various other systemic diseases, Alzheimer's disease, Cardiovascular diseases, Rheumatoid Arthritis, Respiratory diseases, Bacteremia and Cancer. Despite the fact that both Streptococcus mutans and Streptococcus sobrinus are the major etiologic agents of dental caries, S. mutans is more prevalent than S. sobrinus in dental plaque. Early detection of S. mutans and S. sobrinus was carried out from five caries affected dental plaque samples collected from Sri Ramakrishna Dental College and Hospital, Coimbatore, by using semi-quantitative real-time PCR. Specific primers for gtfB and gtfI genes of S. mutans and S. sobrinus respectively were used for the quantification of cariogenic bacteria in the given dental plaque samples. The Biopro Oral Microbiome transport media was prepared to carry dental plaque samples from the hospital to the laboratory. Genomic DNA extraction was done by employing magnetic beads and spin columns provided in the Biopro DNA isolation kit. Various biochemical tests were performed on the bacterial cultures isolated from dental plaque.
半定量实时聚合酶链反应检测人牙菌斑中变形链球菌和隐球菌
龋齿是人类中一种非常普遍的传染病,是由覆盖在牙齿表面的细菌菌斑引起的,是一个严重的公共卫生问题。近年来,牙菌斑的形成与其他各种全身性疾病的发生有关,如阿尔茨海默病、心血管疾病、类风湿关节炎、呼吸系统疾病、菌血症和癌症。尽管变形链球菌和隐球菌都是龋齿的主要病原,但在牙菌斑中,变形链球菌比隐球菌更为普遍。采用半定量实时PCR技术,对哥印拜陀罗摩克里希纳牙科学院和医院采集的5个龋病牙菌斑样本进行了变形链球菌和隐球菌的早期检测。利用变形链球菌gtfB和sobrinus gtfI基因特异性引物对牙菌斑样品中的致龋菌进行定量分析。制备了Biopro口腔微生物组运输培养基,用于将牙菌斑样本从医院运送到实验室。采用Biopro DNA分离试剂盒提供的磁珠和自旋柱进行基因组DNA提取。对从牙菌斑中分离的细菌培养物进行了各种生化试验。
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