Identification of Red Complex Pathogens Group from Chronic Periodontitis Patients in Mosul City

Abstract

This study is concerned with the isolation of red complex pathogens, identifying them by a new molecular method as the first locally used and described procedure and characterizing these pathogens by their phenotypic features and biofilms. Gingival fluids were sampled from chronic periodontitis and inoculated into three types of culture media, Schaedler Anaerobe Agar, Tannerella forsythia (TF) agar, and Trypton Yeast extracts Gelatin Volatile fatty acids and Serum (TYGVS) agar. The different appearing colonies were purified and identified by Loop-Mediated Isothermal Amplification protocol (LAMP) for the detection of red complex species. Bacterial biofilm was estimated in term of mono-and polytypic biofilm by measuring the absorbance of a crystal violet-stained biofilm formed in a microtiter plate. Different forms of colonies appeared at the primary isolation. LAMP method was of a significant value for perfect rapid identification of the target species of extracted DNA or intact cells within half an hour. The three types of red complex pathogens were simultaneously detected in the same gingival fluid sample. They formed mono-and polymicrobial biofilms in a synergistic manner particularly the two intimates P. gingivalis and T. denticola. In conclusion, the updated LAMP molecular protocol was attractive method for the diagnosis of red complex pathogens which showed great morphological variations. P. gingivalis fortified the growth of the other two to establish polymicrobial biofilms in a synergistic manner.

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

“…LAMP is a useful technique in dental practice for the quick identification of periodontopathic bacteria with high sensitivity and specificity. [ 35 40 41 42 ] The current study compared the effectiveness of the LAMP method to that of other molecular techniques, including the conventional PCR and real-time PCR, and established its relevance in dental practice over these commonly used molecular methods.…”

Morphological and molecular identification of a new local strain of Eikenella corrodens isolated from chronic periodontitis

“…The absorbent paper points were transferred directly into tubes containing a modified transport medium. The patients (14 male and 11 female with an age of 19-65 years) proved that they had no systemic disease, were not smokers and had not been taking antibiotics for 3 months before sampling (10).…”