Fever is one of the most common diseases affecting humans, as it results from any disease or development and worsening of the disease for most people with widespread infections in the body. Therefore, this study aimed to evaluate antibiotic resistance genes (CTX-M, Van A and Van B) of Enterococcus faecalis isolated from children with bacteremia by RT-PCR. A total of 200 children was enrolled in the study, 100 children with fever and 100 healthy children (not suffering from any problem); that is, they are a control group for the detection of antibiotic resistance genes (CTX-M, Van A and Van B) of Enterococcus faecalis by RT-PCR. The age of the two groups ranged from one to five years. Four ml of venous blood sample was collected from each child; the venipuncture area was sterilized first with alcohol at a rate of 70%, followed by medical iodine and then sterilized with alcohol again to avoid contamination with skin flora. The blood samples were cultured on media for isolating bacteria. Then, the resistant isolates of E. faecalis to Vancomycin and cefotaxime antibiotics were taken and kept in special nutrient agar media where the DNA of the bacteria was extracted using (Zymogene Extraction kit, Japan). The detection of the exact genes (CTX-M, Van A and Van B) was done using Real-Time PCR technology according to the protocol mentioned by the company (Sacace biotechnology, Italy). The study presented that 40% of children with fever have positive blood cultures compared with 5% in the control group, with a significant difference between the two groups (P<0.001). The study found that 32.5% of bacteremic children were due to S. aureus, 30%, 5%, and 4% were due to E. faecalis, E. coli, P. aeruginosa and Klebsiella spp, respectively, with significant difference (P<0.01). The study showed that 91.67% of E. faecalis isolates were sensitive to Levofloxacin, 83.33% to Amoxiclav, 66.67% to Erythromycin, 58.33% to Amikacin, 50% to Ampicillin, 33.33% to cefotaxime and Ceftriaxone and 25% toward Vancomycin. From 9 isolates resistant to Vancomycin, the study presented that 88.89% of them were observed with Van A gene production as detected by real-time PCR (P<0.001). The study also showed that 77.78% were observed with Van B gene production as detected by real-time PCR (P<0.001). The study revealed that all E. faecalis isolates resistant to cefotaxime and Ceftriaxone were characterized by CTX gene production as detected by real-time PCR (P<0.001). |
- Marston HD, Dixon DM, Knisely JM, Palmore TN, Fauci AS. Antimicrobial resistance. JAMA. 2016;316(11):1193-204.
- Newland JG, Schuster J. Infections in Children, An Issue of Infectious Disease Clinics of North America, E-Book: Elsevier Health Sciences; 2018.
- Friedman G, Stepensky P, Ahmad WA, Masarwa R, Temper V, Oster Y, et al. Enterococcal bacteremia in children with malignancies and following hematopoietic stem cell transplantation: a 15-year single-center experience. Pediatr Infect Dis J. 2020;39(4):318-24.
- Morales-Espinosa R, Delgado G, Espinosa LF, Isselo D, Mendez JL, Rodriguez C, et al. Fingerprint analysis and identification of strains ST309 as a potential high risk clone in a Pseudomonas aeruginosa population isolated from children with bacteremia in Mexico City. Front Microbiol. 2017;8:313.
- Beckman M, Washam MC, DeBurger B, Haslam DB, Courter JD, Andersen H, et al. Reliability of the Verigene system for the identification for Gram-positive Bacteria and detection of antimicrobial resistance markers from children with bacteremia. Diagn Microbiol Infect Dis. 2019;93(3):191-5.
- Dodson DS, Dominguez SR, MacBrayne CE, Williams MC, Parker SK, editors. Vancomycin-nonsusceptible enterococci mediated by vanC at a large children’s hospital: prevalence, susceptibility, and impact on care of enterococcal bacteremia. Open Forum Infectious Diseases; 2020: Oxford University Press US.
- Furuichi M, Furuichi M, Horikoshi Y, Miyairi I. Infectious diseases consultation improves treatment and decreases mortality by enterococcal bacteremia in children. Pediatr Infect Dis J. 2018;37(9):856-60.
- Campbell AJ, Daley DA, Bell JM, Pang S, Coombs GW, Carapetis JR, et al. Progress towards a coordinated, national paediatric antimicrobial resistance surveillance programme: Staphylococcus aureus, enterococcal and Gram-negative bacteraemia in Australia. J Antimicrob Chemother. 2020;75(6):1639-44.
- Kateete DP, Edolu M, Kigozi E, Kisukye J, Baluku H, Mwiine FN, et al. Species, antibiotic susceptibility profiles and van gene frequencies among enterococci isolated from patients at Mulago National Referral Hospital in Kampala, Uganda. BMC Infect Dis. 2019;19(1):1-9.
- Rengaraj R, Mariappan S, Sekar U, Kamalanadhan A. Detection of vancomycin resistance among Enterococcus faecalis and Staphylococcus aureus. Journal of clinical and diagnostic research: JCDR. 2016;10(2):DC04.
- Shokoohizadeh L, Ekrami A, Labibzadeh M, Ali L, Alavi SM. Antimicrobial resistance patterns and virulence factors of enterococci isolates in hospitalized burn patients. BMC Res Notes. 2018;11(1):1-5.
- Hammerum AM, Baig S, Kamel Y, Roer L, Pinholt M, Gumpert H, et al. Emergence of vanA Enterococcus faecium in Denmark, 2005–15. J Antimicrob Chemother. 2017;72(8):2184-90.
- Jahansepas A, Aghazadeh M, Rezaee MA, Hasani A, Sharifi Y, Aghazadeh T, et al. Occurrence of Enterococcus faecalis and Enterococcus faecium in various clinical infections: detection of their drug resistance and virulence determinants. Microb Drug Resist. 2018;24(1):76-82.
- Raza T, Ullah SR, Mehmood K, Andleeb S. Vancomycin resistant Enterococci: A brief review. J Pak Med Assoc. 2018;68(5):768-72.
- Sever JL, Ellenberg JH, Ley AC, Madden DL, Fuccillo DA, Tzan NR, et al. Toxoplasmosis: maternal and pediatric findings in 23,000 pregnancies. Pediatrics. 1988;82(2):181-92.
- Almahdawy OT, Pricop R, Sadik O, Najee H, Pircalabioru GG, Marutescu L, et al. Description of vancomycin resistance genes in Enterococcus sp. clinical strains isolated from Bucharest, Romania. Rom Biotechnol Lett. 2019;24:395-9.
- Guzman Prieto AM, van Schaik W, Rogers MR, Coque TM, Baquero F, Corander J, et al. Global emergence and dissemination of enterococci as nosocomial pathogens: attack of the clones? Front Microbiol. 2016;7:788.
- Hammerum AM, Justesen US, Pinholt M, Roer L, Kaya H, Worning P, et al. Surveillance of vancomycin-resistant enterococci reveals shift in dominating clones and national spread of a vancomycin-variable vanA Enterococcus faecium ST1421-CT1134 clone, Denmark, 2015 to March 2019. Eurosurveillance. 2019;24(34):1900503.
|