This document summarizes a study that used real-time PCR (qPCR) to detect the presence of Entamoeba gingivalis in plaque-induced gingivitis patients. Saliva samples were collected from 50 patients with plaque-induced gingivitis and 50 healthy controls. qPCR analysis found E. gingivalis in 29 of the patient samples (58%) but none of the healthy control samples. No E. gingivalis was detected in the healthy controls. The study demonstrates that qPCR is a more sensitive method for detecting E. gingivalis than traditional diagnostic tools, and finds a higher prevalence of E. gingivalis in patients with plaque-induced gingiv

1. Molecular Detection of Entamoeba gingivalis Using of Real Time (q PCR) technique
in Plague Induced Gingivitis Patients
Prepared by:
Samara Abdulhameed AL-Salihi
University of Babylon/College of Medicine

2. Introduction
Entomoeba. gingivalis is a protozoan endoparasite, living in tartar
and puspockets of the teeth of pyorrhoea infected human beings,
The first definition of Entamoeba gingivalis then called "Amoebea
gengivalis" was made in 1849 from dental plaque tests, referring
to amoebic movement and the development of internal vesicles
(Gros, 1849).E. gingivalis are found on the teeth and gum surface,
gingival pockets near the base of the teeth, in the tonsil crypts and
tonsil tissue sections, calculus and bacterial plaques, gingival
crevicular fluid (GCF), and saliva of patients with
periodontal disease
Real-time PCR used to determine the incidence of E. gingivalis in
healthful gingival pockets (Trim et al., 2011), molecular studies for
detection of E.gingivalis and development

3. Problem of study
Detection of E. gingivalis use RT( qPCR)
technique in plaque-induced
patients with gingivitis and in healthy oral
individuals.

4. Importance and Objectives of
Entamoeba gingivalis
.importance Entamoeba gingivalis in the gingival tissue is an important agent in
occurrence of gingivitis and periodontitis. Other studies, also, consider it an
opportunist that is able to survive in the medium induced by periodontal disease.
.objectives Detection of Entamoeba gingivalis Using of Real Time (q PCR) technique in
Plague Induced Gingivitis Patients in Babylon

5. Methodology of the study
Samples Collection:
The study population involved 100 subjects between (10-
30)years of age ,50 patients (Case Control) who were taken
from patients clinically diagnosed with PIG (plaqe-induced
gingivitis) visiting the dental clinics College of Dentistry,
University of Babylon, Iraq,and specialized dental center in
hilla city, and from 50person who were (Healthy Control),
saliva samples collected in contaniers gaven profile, Number
and patient information babylon university college of dentistry
provided ethical approval for this research, then moved in
specific refrigerator tubes (deep freezing) for molecular
testing in (ASCO leraning center) laboratories Alharithia
Baghdad.

6. Results

7. A. Cycling: E.gingivalis.
Figure (1)Real Time PCR amplification for Entamoeba gingivalis of 50 saliva
samples in PIG disease case group

8. B.Melt: Entamoeba gingivalis
melting curves analysis (Temperatures °C) Real Time PCR of 50 Saliva samples
in Case Group,The melting Temperatures of Entamoeba gingivalis gene starting
from 76.93 °C, 77.90 °C To 78.97 °C
Figure (2) melting curves analysis (Temperatures °C) Real Time PCR of 50
saliva samples Entamoeba .gingivalis

9. Table(1)Saliva real time PCR assay results in PIG patients and
healthy control persons for Entamoeba. gingivalis
P < 0.05
Table (2): Distribution of Entamoeba. gingivalis in PIG patients
according to the sex
%
E. gingivalis
Infected
No
No. Examined
Gender
44
22
30
Male
14
7
20
Female
58
29
50
Total
P value< 0.05
RT(PCR) PIG % Controls %
Nagtive 21 42 50 1
Positive 29 58 0 0
Total 50 100 50 100

10. Conclusion
Using Q(RT- PCR) technique and melting curves analysis
was more sensitive and accurate with rapid results in
diagnosis of Entamoeba. gingivalis with traditional tools for
diagnosis of parasites The finding in this study showed
prevalence of Entamoeba .gingivalis higher with PIG disease
patients 58% .

11. References
1.Maybodi FR, Ardakani AH, Bafghi AF, Ardakani AH, Zafarbakhsh A. The
Effect of NonsurgicalPeriodontal Therapy on Trichomonas Tenax and
Entamoeba Gingivalis in Patients with Chronic Periodontitis. J Dent Shiraz
Univ Med Sci. 2016;17: 171.
2. . Albuquerque RLC Jr, de Melo CM, de Santana WA, Ribeiro JL, Silva FA.
Incidence of Entamoeba gingivalis and Trichomonas tenax in samples of
dental biofilm and saliva from patients with periodontal disease. Rev
Gaْ cha Odontol. 2011; 59:

12. Thanks