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Table of Contents
ORIGINAL ARTICLE
Year : 2021  |  Volume : 11  |  Issue : 1  |  Page : 17-20

Assessment of correlation between presence of red complex bacterias and periodontitis


Department of Periodontics and Oral Implantology, Institute of Dental Sciences, Jammu, Jammu and Kashmir, India

Date of Submission08-Jul-2020
Date of Acceptance06-Jan-2021
Date of Web Publication22-Apr-2021

Correspondence Address:
Dr. Malvika Singh
of Periodontics and Oral Implantology, Institute of Dental Sciences, Jammu, Jammu and Kashmir
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/jid.jid_62_20

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   Abstract 


Context: Oral cavity offers diverse habitats to various species of different microorganisms; some of them under certain conditions cause loss of connective tissue and bone ultimately leading to periodontitis. Once such complex of bacteria often referred to as red complex bacteria consist of Porphyromonas gingivalis, Treponema denticola and Tannerella forsythia have been regarded as the one associated with severe form of periodontitis. With increasing advances in the field of medicine N-benzoyl-DL-arginine-2-naphthylamide (BANA) test, a modern chairside paraclinical method has been designed to detect the presence of one or more such bacteria. Aim: The aim of the study was to detect the presence of red complex bacteria in patients suffering from various forms of periodontitis using BANA test kit. Aims: The aim of this study was to assess the correlation between the presence of red complex bacteria and the severity of periodontal disease. Settings and Design: A total of 150 patients suffering from various forms of periodontitis were selected for the study. Subjects and Methods: These were divided into three groups – Group I: mild periodontitis, Group II: moderate periodontitis, and Group III: severe periodontitis. Microbial parameters were recorded using BANA test. Statistical Analysis Used: Mann–Whitney test and Pearson's Chi-square test were used for statistical analysis. Results: Group III had significantly greater percentage of BANA-positive sites, followed by Group II and Group I. Conclusions: There is a positive correlation between the presence of red complex bacteria and the severity of periodontitis.

Keywords: Chronic periodontitis, N-benzoyl-DL-arginine-2-naphthylamide, oral microbiota, red complex bacteria


How to cite this article:
Singh M. Assessment of correlation between presence of red complex bacterias and periodontitis. J Interdiscip Dentistry 2021;11:17-20

How to cite this URL:
Singh M. Assessment of correlation between presence of red complex bacterias and periodontitis. J Interdiscip Dentistry [serial online] 2021 [cited 2021 Jul 25];11:17-20. Available from: https://www.jidonline.com/text.asp?2021/11/1/17/314182




   Clinical Relevance to Interdisciplinary Dentistry Top


Detection of red complex bacteria in early stages not only reduces the progression of periodontal disease but their mere detection can reduce the development of endodontic problems and can reduce the chances of development of any prosthodontic failure.


   Introduction Top


Oral cavity offers diverse habitats wherein different species of microorganisms can prosper and an aggregation of which is referred to as dental plaque. Microbes in dental plaque flourish in niche to adhere to the tooth surfaces and multiply in shielded environment such as periodontal pockets and tooth crevices. As proposed by Socransky et al., most of the pathogenic of all complexes comprised Porphyromonas gingivalis (Pg), Tannerella forsythia, and Treponema denticola (Td) also collectively known as red complex. These red complex bacteria have shown to be most important periodontal pathogens in causing periodontal disease[1] by production of various trypsin-like enzymes that degrade intercellular matrix of periodontal tissues.[2] With advances in technology, the diagnostic tools these days are based on enzymes diagnostic markers to identify specific periodontopathic bacteria which helps to provide preventive and therapeutic measures toward disease control. N-benzoyl-DL-arginine-2-naphthylamide (BANA) enzymatic test is a rapid and reliable chairside diagnostic test, which can be performed in about 15-min time and can give information about the presence of three of the putative pathogens in subgingival plaque samples. Loesche et al. studied a strong relationship between a BANA-positive reaction and high levels of plaque spirochetes and proposed that BANA test detects the presence of periodontal pathogens, thus making it as marker for disease activity while monitoring the activity of periodontopathogen bacteria too.[3]

The aim of this study was to assess the correlation between red complex bacteria and severity of periodontitis using BANA test kit.


   Subjects and Methods Top


The present study was carried out in the Department of Periodontology and Oral Implantology of our institute. Patients who fulfilled the following criteria were included and excluded from the study:

Inclusion criteria

  1. Patients suffering from different forms of periodontitis belonging an age group of 35–75 years
  2. Patients having bilateral pockets of 5–7 mm in molars.


Exclusion criteria

  1. Those patients who presented themselves with any systemic disease
  2. Patients undergoing any local or systemic antimicrobial and anti-inflammatory therapy for the past 6 months
  3. Patients on periodontal therapy other than standard prophylaxis during previous 6 months
  4. Pregnant, lactating women, tobacco chewers, and alcoholics.


The study was approved from the Institute Ethical Committee before the start of the study.

Study design

A simple randomized, clinical study was conducted to check the presence of red complex bacteria in periodontitis patients with various degrees of severity. A total of 150 patients with 300 sites were randomly divided into the following three groups based on case definition and classification given by the Prevention and American Academy of Periodontology [Table 1][4],[5] into:
Table 1: Prevention (case definition and classification) and American Academy of Periodontology

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  1. Group I (50 patients and 100 sites): Patients suffering from mild periodontitis
  2. Group II (50 patients and 100 sites): Patients suffering from moderate periodontitis
  3. Group III (50 patients and 100 sites): Patients suffering from severe periodontitis.


The nature and design of the clinical study were explained and informed consent was obtained from all the participants and BANA chair side test was performed.

Procedure

Subgingival plaque sample from the periodontal pocket of each patient was obtained using curette and applied on to the raised reagent matrix affixed to the lower portion of the test strip. The upper test strip was moistened with distilled water using a cotton swab. BANA test strip was folded at the given crease mark so that the lower and upper reagent strips were met. It was then placed into either of the slots on the top of the processor. The heating element of the processor started automatically when strip was inserted into the bottom of the slot, as indicated by the flashing light. The BANA test color was indicated when the indicator light went off and the bell rang. BANA strip was removed from the processor, and the lower reagent strip was then discarded to prevent the contamination of it further. The upper reagent strip was examined for the presence of blue color. If a blue color was a detected, the site was marked as either weak positive or positive. The darkness of blue color ranged from deep blue (for patients with severe periodontitis) to pale blue (for patients with moderate periodontitis). However, as done for the patients of gingivitis, the color ranged from light yellow to salmon color [Figure 1], [Figure 2], [Figure 3] depending on the severity of gingivitis indicating the test to be weak negative as red complex bacteria were not detected in the same group. Recording was done for each sampled site as negative, weak positive, or positive.
Figure 1: BANA test for patients suffering from mild periodontitis

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Figure 2: BANA test for patients suffering from moderate periodontitis

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Figure 3: BANA test for patients suffering from severe periodontitis

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   Results Top


All patients (100 males and 50 females with a mean age of 55 ± 5 years) completed the study. Recordings were carried out on the 1st day before treatment. All recordings were subjected for statistical analysis using Mann–Whitney and Pearson's Chi-square tests. Microbiological assessment between the groups showed highly significant [Table 2]. However, it showed that BANA is sensitive but has low specificity as the levels of individual bacteria could not be judged after performing BANA test.
Table 2: Color change in patients using N-benzoyl-DL-arginine-2-naphthylamide test

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   Discussion Top


Periodontitis, an infectious disease caused by bacteria, brings about destructive changes leading to loss of bone and connective tissue attachment. Several oral bacteria such as the Gram-negative anaerobic rods Pg, T. forsythia, and Td have been considered as a major pathogen in periodontitis. These bacteria were most crucial for the progression of this disease, and thus, their detection of these organisms provides essential information about the severity of periodontitis. Various systems for accurate assessment of the periodontal status prior and during the periodontal treatment based on the presence of oral pathogens are available today in the market. Once such among them is microbial-enzymatic N benzoyl-DL-arginine-2-napthylamide or BANA test kit. It is one of the modern alternatives to bacterial cultures and detects the presence of three periodontal pathogens in the subgingival plaque. The study was conducted in 150 patients to evaluate the association of red complex bacteria with severity of periodontal disease in patients suffering from chronic periodontitis. The microbiological assessment was performed using a chairside BANA test showing specific microbiota. Pg, T. forsythia, and Td species share a common enzymatic profile and have a trypsin-like enzyme in common. This activity of enzyme can be measured with the hydrolysis of the colorless substrate BANA; hence, all the three BANA-positive species are frequently cited as potential periodontal pathogens.[6] In our study, we found out the presence of these pathogens in all subgingival pockets and their number increased with the severity of the periodontal disease which was confirmed by performing BANA test. Subgingival plaque sample belonging to the patient of mild periodontitis gave salmon pink color on the BANA strip, which stained pale blue and deep blue in patients suffering from moderate to severe periodontitis, respectively. Thus, a positive correlation between the presence of red complex bacteria and the severity of periodontitis was found. The percentage of BANA-positive sites was significantly higher for Group III, followed by Group II and Group I [Graph 1], [Graph 2], [Graph 3]. This was attributed to the presence of more red complex bacteria which clinically showed more periodontal destruction in severe periodontitis as compared to moderate and mild forms of periodontitis.



The advantages of using BANA test were that BANA test could be used as a reliable indicator of BANA-positive species in dental plaque. It could also be used for detection of periodontitis for epidemiological purposes and could be used as objective means of determining diseased sites, requiring some forms of periodontal treatment. However, the disadvantage was that this test was highly sensitive but not specific, i.e., it could detect the presence of red complex bacteria as a group but could specifically detect them as individual bacteria.


   Conclusions Top


There is a positive correlation between the presence of red complex bacteria and the severity of periodontitis, and BANA test can be used as a reliable indicator of BANA-positive species in dental plaque.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
   References Top

1.
Socransky SS, Haffajee AD, Cugini MA, Smith C, Kent RL Jr. Microbial complexes in subgingival plaque. J Clin Periodontol 1998;25:134-44.  Back to cited text no. 1
    
2.
Slots J, Bragd L, Wikström M, Dahlén G. The occurrence of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Bacteroides intermedius in destructive periodontal disease in adults. J Clin Periodontol 1986;13:570-7.  Back to cited text no. 2
    
3.
Loesche WJ, Lopatin DE, Giordano J, Alcoforado G, Hujoel P. BANA as accurate as DNA testing for periodontal pathogens. J Clin Microbiol 1992;30:427-33.  Back to cited text no. 3
    
4.
Page RC, Eke PI. Case definitions for use in population-based surveillance of periodontitis. J Periodontol 2007;78 Suppl 7S:1387-99.  Back to cited text no. 4
    
5.
Eke PI, Page RC, Wei L, Thornton-Evans G, Genco RJ. Update of the case definitions for population-based surveillance of periodontitis. J Periodontol 2012;83:1449-54.  Back to cited text no. 5
    
6.
Amalfitano J, de Filippo AB, Bretz WA, Loesche WJ. The effects of incubation length and temperature on the specificity and sensitivity of the BANA (N-benzoyl-DL-arginine-naphthylamide) test. J Periodontol 1993;64:848-52.  Back to cited text no. 6
    


    Figures

  [Figure 1], [Figure 2], [Figure 3]
 
 
    Tables

  [Table 1], [Table 2]



 

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