Microbial Profiling in Pockets Related to Chronic Periodontitis Patients Using 16s RNA Metagenomics Sequencing

Sponsor

Ajman University (Other)

Overall Status

Completed

CT.gov ID

NCT04425343

Collaborator

(none)

Enrollment

Location

Arms

22.9

Actual Duration (Months)

3.5

Patients Per Site Per Month

Study Details

Study Description

Brief Summary

The study was developed in compliance with the Helsinki Declaration on medical research involving the Ethics Committee at Ajman University (2017-A-DN-04). Informed consent from all the participants was obtained before participation in the study. The participants were systemically healthy with no history of antibiotics for the past three months. Patients recruited for the study were diagnosed as Stage II Generalized periodontitis according to the classification from the 2017 world workshop on the "classification of periodontal and peri-implant disease and conditions". Periodontal status indicating the severity of interdental clinical attachment loss of 3-4mm, radiographic bone loss between 15%-33% and with no tooth loss were included. The complexity of periodontitis with a maximum probing depth of ≤ 5mm with horizontal bone loss and having an extent and distribution with >30% teeth involved were included in the study. A total of 80 plaque samples were collected with the clinical characteristics of the patient comprising of age between 25-39 years, 36 females and 44 males. The subgingival plaque was collected using a sterile curette from the buccal aspect of maxillary molars and lingual aspect of mandibular incisors, in a vial containing 200µl of Buffer CL.

Condition or Disease	Intervention/Treatment	Phase
Periodontal Diseases	Genetic: 16s RNA Metagenomics sequencing	N/A

Detailed Description

The collected plaque samples were then freshly prepared for the DNA isolation using ABIOpure TM Total DNA (version 2.0) (Cat No: M501DP100) according to the manufacturer's instruction. All the samples were assessed for DNA quantification using spectrophotometry and further quantified using the fluorometric method. This was performed using DeNovix DS-11 FX (DeNovix). Further to the DNA quantification, assessment of DNA integrity was resolved on a 0.8% agarose gel with ethidium bromide. For NGS library preparation, all the samples underwent PCR amplification of the 16S rRNA gene in the isolated bacterial DNA. The primers used had targeted in the V3-V4 region of the 16S rRNA gene. The full length primer sequences, using standard IUPAC nucleotide nomenclature, to follow the protocol targeting this region are:

16SAmpliconPCRForwardPrimer=5' TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGCCTACGGGNGGCWGCAG 16SAmpliconPCRReversePrimer=5' GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGGACTACHVGGGTATCTAATCC Gel electrophoresis method was used for the size selection and the bioinformatics pipeline used for processing microbiome 16S sequence data was QIIME

Study Design

Study Type:

Interventional

Actual Enrollment :

80 participants

Allocation:

Randomized

Intervention Model:

Single Group Assignment

Intervention Model Description:

. Patients recruited for the study were diagnosed as Stage II Generalized periodontitis according to the classification from the 2017 world workshop on the "classification of periodontal and peri-implant disease and conditions". Periodontal status indicating the severity of interdental clinical attachment loss of 3-4mm, radiographic bone loss between 15%-33% and with no tooth loss were included. The complexity of periodontitis with a maximum probing depth of ≤ 5mm with horizontal bone loss and having an extent and distribution with >30% teeth involved were included in the study.. Patients recruited for the study were diagnosed as Stage II Generalized periodontitis according to the classification from the 2017 world workshop on the "classification of periodontal and peri-implant disease and conditions". Periodontal status indicating the severity of interdental clinical attachment loss of 3-4mm, radiographic bone loss between 15%-33% and with no tooth loss were included. The complexity of periodontitis with a maximum probing depth of ≤ 5mm with horizontal bone loss and having an extent and distribution with >30% teeth involved were included in the study.

Masking:

Single (Investigator)

Masking Description:

. A total of 80 plaque samples were collected with the clinical characteristics of the patient comprising of age between 25-39 years, 36 females and 44 males

Primary Purpose:

Health Services Research

Official Title:

Microbial Profiling in Pockets Related to Chronic Periodontitis Patients in UAE Population Using 16s RNA Metagenomics Sequencing

Actual Study Start Date :

Nov 10, 2017

Actual Primary Completion Date :

Sep 15, 2018

Actual Study Completion Date :

Oct 7, 2019

Arms and Interventions

Arm	Intervention/Treatment
Experimental: Periodontitis, Adult Plaque samples were taken from subgingival pocket and send to the lab for metagenomic analysis	Genetic: 16s RNA Metagenomics sequencing The subgingival plaque was collected using a sterile curette from the buccal aspect of maxillary molars and lingual aspect of mandibular incisors, in a vial containing 200µl of Buffer CL. Other Names: DNA isolation
Experimental: Metgenomic analysis Analysis for whole bacterial count	Genetic: 16s RNA Metagenomics sequencing The subgingival plaque was collected using a sterile curette from the buccal aspect of maxillary molars and lingual aspect of mandibular incisors, in a vial containing 200µl of Buffer CL. Other Names: DNA isolation

Arm

Intervention/Treatment

Experimental: Periodontitis, Adult

Plaque samples were taken from subgingival pocket and send to the lab for metagenomic analysis

Genetic: 16s RNA Metagenomics sequencing

The subgingival plaque was collected using a sterile curette from the buccal aspect of maxillary molars and lingual aspect of mandibular incisors, in a vial containing 200µl of Buffer CL.

Other Names:

DNA isolation

Experimental: Metgenomic analysis

Analysis for whole bacterial count

Genetic: 16s RNA Metagenomics sequencing

The subgingival plaque was collected using a sterile curette from the buccal aspect of maxillary molars and lingual aspect of mandibular incisors, in a vial containing 200µl of Buffer CL.

Other Names:

DNA isolation

Outcome Measures

Primary Outcome Measures

Microbial Taxonomical Composition [From Baseline to 3 months]
The composition was determined using subgingival plaque samples and performing 16s metagenomic sequencing

Eligibility Criteria

Criteria

Ages Eligible for Study:

18 Years to 60 Years

Sexes Eligible for Study:

All

Accepts Healthy Volunteers:

Yes

Inclusion Criteria:

Patients recruited for the study were diagnosed as Stage II Generalized periodontitis
Medically fit

Exclusion Criteria:

Patients having gingivitis or stage III periodontitis
Medically unfit

Contacts and Locations

Locations

	Site	City	State	Country	Postal Code
1	Ajman University of Science and Technology	Ajman		United Arab Emirates	009716

Sponsors and Collaborators

Ajman University

Investigators

Principal Investigator: Sudhir Varma, MDS, Assistant Professor

Study Documents (Full-Text)

None provided.

More Information

Publications

None provided.

Responsible Party:

sudhir rama varma, Assistant Professor, Ajman University

ClinicalTrials.gov Identifier:

NCT04425343

Other Study ID Numbers:

F-H-17-11-03

First Posted:

Jun 11, 2020

Last Update Posted:

Jun 11, 2020

Last Verified:

Jun 1, 2020

Individual Participant Data (IPD) Sharing Statement:

Plan to Share IPD:

Studies a U.S. FDA-regulated Drug Product:

Studies a U.S. FDA-regulated Device Product:

Keywords provided by sudhir rama varma, Assistant Professor, Ajman University

metagenomics

16S

Periodontitis, Adult

Additional relevant MeSH terms:

Periodontitis

Periodontal Diseases

Chronic Periodontitis

Study Results

No Results Posted as of Jun 11, 2020