Molecular Investigation of Enterococcal Surface protein (esp)gene of Enterococcus faecalis isolated from endodontic patients
Abstract
Enterococci are generally considered transient components of oral bacteria that may be a reason for several oral and systemic infections, particularly, those related to dental root canal infections. Aims: The purpose of the current study was to examine the occurrence of Enterococcal Surface protein esp in Enerococcus faecalis which is isolated from infected root canals. Forty samples were collected from endodontic patients who attended the Conservative Treatment Department in the College of Dentistry/Mosul University/Dental Teaching Hospital. Materials and Methods: All samples were examined traditionally by using HiCrom TM Enterococcus faecium Agar base medium and biochemical tests. 16srRNA sequencing was performed to confirm their identity by using the polymerase chain reaction technique. Then all Enterococcus faecalis isolates were examined for the existence of esp gene coding for enterococcal surface protein by using PCR assay. Results: From 40 clinical samples that were obtained, 31 isolates were recognized as E. faecalis by traditional methods, unexpectedly other non-enterococci genera were also grown on HiCrom TM Enterococcus faecium Agar base medium. The PCR products for the sequence-specific primers obtained from the full-length of 16S rRNA gene sequence which belongs to E. faecalis and the PCR products for specific primer of esp genes created bands at the position of 138bp and 932 bp on the agarose gel respectively. This gene which is correlating with the aggregation of this bacteria on the canal walls was detected in high proportion (91%) from the isolates. Conclusions: PCR assay provides an accurate, rapid and more sensitive detection of E. faecalis. A positive correlation has been found between esp gene and enterococcal infections in root canals.
References
1. Mustafa EA, Hamdoon SM, Shehab EY. Molecular detection and identification of Enterococcus faecium isolated from dental root canals. Iraqi Journal of Science. 2021 May 30:1447-51.
2. Akhondnezhad M, Bakhti M, Nasrolahei M, Shabankhani B, Goli HR. Molecular detection of Enterococcal surface protein (esp) gene in Enterococcus faecalis isolated from dental calculus of patients in Sari, Iran. Research in Molecular Medicine. 2017 Aug 10;5(3):21-5.
3. Kouidhi B, Zmantar T, Mahdouani K, Hentati H, Bakhrouf A. Antibiotic resistance and adhesion properties of oral Enterococci associated to dental caries. BMC microbiology. 2011 Dec;11(1):1-7.
4. Siqueira JF, Rôças IN, Ricucci D. Microbial and nonmicrobial etiologies of endodontic diseases. Dalam: Ingle JI, Rotstein I. Endodontics 7th ed. Raleigh, North Carolina: PMPH-USA Ltd. 2019:85-109.
5. Wang L, Dong M, Zheng J, Song Q, Yin W, Li J, Niu W. Relationship of biofilm formation and gelE gene expression in Enterococcus faecalis recovered from root canals in patients requiring endodontic retreatment. Journal of endodontics. 2011 May 1;37(5):631-6.
6. Fallico L, Boldrin C, Grossato A, Franchin E, De Canale E, Tommasini T, Parisi SG, Manganelli R, Palu G. Molecular epidemiology of Enterococcus faecium isolates from an Italian hospital. Infection. 2011 Apr;39:127-33.
7. Wu H, Moser C, Wang HZ, Høiby N, Song ZJ. Strategies for combating bacterial biofilm infections. International journal of oral science. 2015 Mar;7(1):1-7.
8. Alnasra NA, Badran EF, Dajan N, Shehabi AA. Antimicrobial susceptibility and virulence factors of Enterococci colonizing intestinal tract of infants. The International Arabic Journal of Antimicrobial Agents. 2016 May 3;6(1).
9. Taglialegna A, Matilla-Cuenca L, Dorado-Morales P, Navarro S, Ventura S, Garnett JA, Lasa I, Valle J. The biofilm-associated surface protein Esp of Enterococcus faecalis forms amyloid-like fibers. npj Biofilms and Microbiomes. 2020 Mar 27;6(1):15.
10. Abdulrazzaq R, Faisal R. Efficiency of hichrome Enterococcus faecium agar in the isolation of Enterococcus spp. and other associated bacterial genera from water. Journal of Life and Bio Sciences Research. 2022 Jan 26;3(01):01-6.
11. Gary P, Hall G. Koneman’s Color Atlas and Textbook of Diagnostic Microbiology Color: Wolters Kluwer Health,2017.
12. Sedgley CM, Molander A, Flannagan SE, Nagel AC, Appelbe OK, Clewell DB, Dahlén G. Virulence, phenotype and genotype characteristics of endodontic Enterococcus spp. Oral microbiology and immunology. 2005 Feb;20(1):10-9.
13. Khalid I, Nayyef NS, Merkhan MM. A Taxonomic Study comparing the two types of Medicinal Leeches available in Iraq. Research Journal of Pharmacy and Technology. 2022 Mar 1;15(3):1119-22.
14. Al-Askary RA. Preparation and Evaluation of Glutamic Acid - Calcium Oxide Based Endodontic Sealer.Ph.D.thesis.Collage of Dentistry. Mosul University.Iraq.
15. Komiyama EY, Lepesqueur LS, Yassuda CG, Samaranayake LP, Parahitiyawa NB, Balducci I, Koga-Ito CY. Enterococcus species in the oral cavity: prevalence, virulence factors and antimicrobial susceptibility. PloS one. 2016 Sep 15;11(9):e0163001.
16. Durgesh BH, Alkheraif AA, Malash AM, Hashem MI, Assery MK, Al Asmari M, Durgesh P. Genotypic characterization of species isolated from the oral cavity and their pattern of antibiotic susceptibility. Asian biomedicine. 2016 Feb 1;10(1):49-53.
17. Weng PL, Ramli R, Hamat RA. Antibiotic susceptibility patterns, biofilm formation and esp gene among clinical Enterococci: is there any association?. International journal of environmental research and public health. 2019 Sep;16(18):3439.
18. Angeletti S, Lorino G, Gherardi G, Battistoni F, De Cesaris M, Dicuonzo G. Routine molecular identification of enterococci by gene-specific PCR and 16S ribosomal DNA sequencing. Journal of clinical microbiology. 2001 Feb 1;39(2):794-7.
19. d'Azevedo PA, Santiago KA, Furtado GH, Xavier DB, Pignatari AC, Titze-de-Almeida R. Rapid detection of vancomycin-resistant enterococci (VRE) in rectal samples from patients admitted to intensive care units. Brazilian Journal of Infectious Diseases. 2009;13:289-93.