Evaluation of loop-mediated isothermal amplification method for efficient detection of the periodontopathic bacteria Porphyromonas gingivalis.

Q2 Dentistry
Meenaz N Sangolli, Manohar S Kugaji, Suman Kumar Ray, Kishore G Bhat
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引用次数: 0

Abstract

Background: Periodontitis is a multifactorial, polymicrobial oral inflammatory illness brought on by oral pathogens. Porphyromonas gingivalis is a Gram-negative, obligatory anaerobic, black-pigmented coccobacillus and is regarded as a primary etiological factor in the progression of periodontitis. Rapid, highly senstitive and specific detection methods are emerging. The present study aimed to evaluate the loop-mediated isothermal amplification (LAMP) technique for efficiently detecting P. gingivalis from subgingival plaque samples of chronic periodontitis patients.

Materials and methods: This study included 50 subgingival plaque samples from patients suffering from chronic periodontitis. The DNA (Deoxyribonucleic acid) was extracted by the "modified proteinase K" method. A set of six primers, targeting the pepO gene of P. gingivalis, was used for conducting LAMP. The amplification was visualized by naked-eye detection and agarose electrophoresis. Conventional polymerase chain reaction (PCR) and real-time qantitative PCR (qPCR) were carried out by targeting the 16SrRNA (16S ribosomal ribonucleic acid) gene of P. gingivalis.

Results: The results showed that LAMP detected P. gingivalis in 40 out of 50 samples (80%). Whereas, qPCR and conventional PCR technique detected P. gingivalis in 38 (76%) and 33 (66%) samples respectively. The sensitivity and specificity of the LAMP method were 94.87% and 90.90%, respectively. With qPCR, the sensitivity and specificity were found to be 92.30% and 81.81%, respectively, whereas, with conventional PCR, it was found to be 76.92% and 72.72%, respectively.

Conclusion: LAMP is an efficient technique for quick, accurate, and reliable identification of P. gingivalis from subgingival plaque samples. The technique needs to be validated analytically, and further studies can be conducted by taking saliva and/or gingival crevicular fluid samples from periodontitis patients.

评估环介导等温扩增法在有效检测牙周病病菌牙龈卟啉单胞菌方面的应用。
背景:牙周炎是一种由口腔病原体引起的多因素、多微生物口腔炎症性疾病。牙龈卟啉单胞菌(Porphyromonas gingivalis)是一种革兰氏阴性、必须厌氧的黑色色素球菌,被认为是牙周炎恶化的主要致病因素。快速、高灵敏度和特异性的检测方法不断涌现。本研究旨在评估环介导等温扩增(LAMP)技术从慢性牙周炎患者龈下牙菌斑样本中有效检测牙龈脓毒性球菌的能力:本研究包括 50 份来自慢性牙周炎患者的龈下牙菌斑样本。采用 "改良蛋白酶 K "法提取 DNA(脱氧核糖核酸)。使用针对牙龈脓疱病菌 pepO 基因的六组引物进行 LAMP 扩增。扩增结果通过肉眼检测和琼脂糖电泳显现。以牙龈脓杆菌的 16SrRNA (16S 核糖体核糖核酸)基因为靶标,进行常规聚合酶链反应(PCR)和实时定量 PCR(qPCR):结果显示,50 份样本中有 40 份(80%)通过 LAMP 检测出了牙龈脓胞。而 qPCR 和传统 PCR 技术分别在 38 个样本(76%)和 33 个样本(66%)中检测到了牙龈杆菌。LAMP 方法的灵敏度和特异性分别为 94.87% 和 90.90%。qPCR 的灵敏度和特异性分别为 92.30% 和 81.81%,而传统 PCR 的灵敏度和特异性分别为 76.92% 和 72.72%:结论:LAMP 是一种从龈下牙菌斑样本中快速、准确、可靠地鉴定牙龈脓毒性杆菌的有效技术。该技术还需要进行分析验证,并可通过采集牙周炎患者的唾液和/或龈沟液样本进行进一步研究。
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来源期刊
CiteScore
1.70
自引率
0.00%
发文量
87
审稿时长
44 weeks
期刊介绍: The Journal of Indian Society of Periodontology publishes original scientific articles to support practice , education and research in the dental specialty of periodontology and oral implantology. Journal of Indian Society of Periodontology (JISP), is the official publication of the Society and is managed and brought out by the Editor of the society. The journal is published Bimonthly with special issues being brought out for specific occasions. The ISP had a bulletin as its publication for a large number of years and was enhanced as a Journal a few years ago
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