Absolute quantitation of oral bacteria involved in oral cancer by real-time PCR

India carries approximately 34% of the global oral cancer burden. Along with tobacco chewing habit, there are other factors that are being studied globally like the involvement of bacteria in oral cancer. NGS studies have identified few bacterial species and the changes in their abundance in healthy...

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Main Authors: Shriya Sawant, Jinesh Dugad, Deepak Parikh, Harinder Singh
Format: Article
Language:English
Published: Elsevier 2021-03-01
Series:Medicine in Microecology
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2590097821000021
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author Shriya Sawant
Jinesh Dugad
Deepak Parikh
Harinder Singh
author_facet Shriya Sawant
Jinesh Dugad
Deepak Parikh
Harinder Singh
author_sort Shriya Sawant
collection DOAJ
description India carries approximately 34% of the global oral cancer burden. Along with tobacco chewing habit, there are other factors that are being studied globally like the involvement of bacteria in oral cancer. NGS studies have identified few bacterial species and the changes in their abundance in healthy and diseased populations based on 16S rRNA metagenomics. The present study shows a method for absolute bacterial quantification from oral cavity rinse samples. The method is a real-time qPCR assay and based on the fact that certain genes are present in one copy per cell (rpoB gene) and we can correlate the copy numbers of these genes with cell numbers in a sample. This method is more accurate than the NGS 16S rRNA gene-based approach which is multicopy gene. Linear correlation between qPCR assay and cell numbers of a model system was established. Consequently, the assay was performed on oral rinse samples of oral cancer/tobacco chewers and healthy subjects to quantitate significant oral bacterial species. The obtained bacterial quantification correlated well with the previous reports. The developed qPCR method is an efficient, faster and resource-friendly method and can be used to quantify bacterial population in cancer/diseased subjects, and can have application in determining the susceptibility of an individual towards a specific disease.
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spelling doaj.art-9f8de04bfcdc402dace6f032cc38c4ba2022-12-21T22:31:43ZengElsevierMedicine in Microecology2590-09782021-03-017100034Absolute quantitation of oral bacteria involved in oral cancer by real-time PCRShriya Sawant0Jinesh Dugad1Deepak Parikh2Harinder Singh3Department of Biological Sciences, Sunandan Divatia School of Science, NMIMS University, Vile Parle (West), Mumbai, IndiaSomaiya Ayurvihar -Asian Cancer Institute, Off Eastern Express Highway, Behind Everard Nagar, Somaiya Ayurvihar, Sion (East), Mumbai, IndiaSomaiya Ayurvihar -Asian Cancer Institute, Off Eastern Express Highway, Behind Everard Nagar, Somaiya Ayurvihar, Sion (East), Mumbai, IndiaDepartment of Biological Sciences, Sunandan Divatia School of Science, NMIMS University, Vile Parle (West), Mumbai, India; Corresponding author.India carries approximately 34% of the global oral cancer burden. Along with tobacco chewing habit, there are other factors that are being studied globally like the involvement of bacteria in oral cancer. NGS studies have identified few bacterial species and the changes in their abundance in healthy and diseased populations based on 16S rRNA metagenomics. The present study shows a method for absolute bacterial quantification from oral cavity rinse samples. The method is a real-time qPCR assay and based on the fact that certain genes are present in one copy per cell (rpoB gene) and we can correlate the copy numbers of these genes with cell numbers in a sample. This method is more accurate than the NGS 16S rRNA gene-based approach which is multicopy gene. Linear correlation between qPCR assay and cell numbers of a model system was established. Consequently, the assay was performed on oral rinse samples of oral cancer/tobacco chewers and healthy subjects to quantitate significant oral bacterial species. The obtained bacterial quantification correlated well with the previous reports. The developed qPCR method is an efficient, faster and resource-friendly method and can be used to quantify bacterial population in cancer/diseased subjects, and can have application in determining the susceptibility of an individual towards a specific disease.http://www.sciencedirect.com/science/article/pii/S2590097821000021Realtime-PCRAbsolute quantificationSpecificityOral cancerTobacco chewing
spellingShingle Shriya Sawant
Jinesh Dugad
Deepak Parikh
Harinder Singh
Absolute quantitation of oral bacteria involved in oral cancer by real-time PCR
Medicine in Microecology
Realtime-PCR
Absolute quantification
Specificity
Oral cancer
Tobacco chewing
title Absolute quantitation of oral bacteria involved in oral cancer by real-time PCR
title_full Absolute quantitation of oral bacteria involved in oral cancer by real-time PCR
title_fullStr Absolute quantitation of oral bacteria involved in oral cancer by real-time PCR
title_full_unstemmed Absolute quantitation of oral bacteria involved in oral cancer by real-time PCR
title_short Absolute quantitation of oral bacteria involved in oral cancer by real-time PCR
title_sort absolute quantitation of oral bacteria involved in oral cancer by real time pcr
topic Realtime-PCR
Absolute quantification
Specificity
Oral cancer
Tobacco chewing
url http://www.sciencedirect.com/science/article/pii/S2590097821000021
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