Molecular Study of Accessory-Gene-Regulator in Staphylococcus aureus Isolated from Sepsis in Pediatric Patients.

BACKGROUND: Pediatric sepsis due to Staphylococcus aureus (S. aureus) is associated with high morbidity and mortality. Accessory-Gene-Regulator (agr) has a role in the pathogenesis of S. aureus through controlling and regulating the expression of virulence genes. Therefore, the aim of the present study was to investigate the prevalence of genotypes of the agr system in S. aureus isolated from children with sepsis and to assess their relationship to biofilm formation and antibiotic resistance. METHODS: The study was a retrograde cross-sectional study that included 131 children with health care associated sepsis due to S. aureus. The isolated S. aureus was investigated for their ability to form biofilm by microplate method, antibiotic susceptibility pattern by disc diffusion method, and molecular determination of agr genotypes by polymerase chain reaction (PCR). RESULTS: Methicillin resistant S. aureus (MRSA) was defined by resistance to cefoxitin antibiotic disc in 70 (53.4%) of the isolates and biofilm formation was positive in 67 (58%) of the isolates. Molecular study of the agr genes revealed that 54 (41.2%), 40 (30.5%), 27 (20.6%), and 10 (7.5%) of the studied isolates had agr I, agr II, agr III, and agr IV, respectively. In comparison between MRSA and methicillin sensitive S. aureus (MSSA), there was a signif-icant increase in biofilm formation among MRSA (65.7%, p = 0.01) compared to MSSA (34.3%) and an increase in agr genotype I among MRSA (68.6%, p = 0.001) compared to agr I in MSSA (9.8%). There was a significant association with the presence of a central venous catheter (51.4%, p = 0.001) and urinary tract catheter (81.4%, p = 0.001) in children with MRSA compared to children with MSSA (21.3%, OR = 3.9, 95% CI = 1.8 - 8.5 and 36.1%, OR = 7.8, 95% CI 3.5 - 17.3, respectively). CONCLUSIONS: There was an increase in the biofilm formation among S. aureus isolated from pediatric patients with sepsis with a significant increase in MRSA. The agr group I was the main agr gene among the isolated S. aureus. Moreover, agr I was the predominant gene in MRSA isolates and was significantly associated with biofilm formation.

Diagnostic Value of Multiplex Polymerase Chain Reaction in Detection of Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia from Sepsis in Pediatrics.

BACKGROUND: Proper identification of the causative organism in pediatric sepsis is crucial for early diagnosis and prevention of septic shock and organ failure. OBJECTIVES: The aim of the present study was to evaluate the multiplex polymerase chain reaction (PCR) for detection of Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia from positive blood cultures for these pathogens isolated from children with hospital- acquired sepsis compared to the conventional biochemical reactions for identification of these organisms. METHODS: This study was a cross-sectional study performed on 100 isolates from pediatric blood cultures, including Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. The study also included 100 isolates of Escherichia coli as a negative control. All isolates were identified by API 20NE and the multiplex PCR with primers specific to the 3 tested bacteria. RESULTS: Multiplex PCR was positive in 96% of isolates and 4 isolates had negative results. Falsepositive results were reported with three E. coli strains. Multiplex PCR identified all the isolates of Acinetobacter baumannii, 29 isolates of Pseudomonas aeruginosa and 27 isolates of Stenotrophomonas maltophilia. The diagnostic value of the multiplex PCR compared to the biochemical identification revealed sensitivity 96.04%, specificity 96.9%, positive predictive value 97.00%, negative predictive value 96.00% and accuracy 96.50%. CONCLUSION: The present study highlights the diagnostic value of multiplex PCR to identify Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia from positive blood cultures. Multiplex PCR was sensitive, specific and accurate. The accuracy differs according to the organisms with 100% accuracy for Acinetobacter baumannii.

Molecular characterization of Staphylococcus aureus isolated from hospital acquired sepsis in pediatrics, relation to antibiotics, resistance and virulence genes.

INTRODUCTION: The objective of this study was to determine the prevalence of antibiotic resistance genes mecA, vanA, B, C and virulence genes Panton-Valentine Leucocidin (PVL) and fibronectin-binding protein (fnBPA) among S. aureus isolates from hospital-acquired sepsis from pediatric intensive care units. METHODS: The study was a retrospective cross-sectional study, including 250 unique isolates of S. aureus obtained from pediatric patients with hospital-acquired sepsis. The isolates were subjected to study of antibiotic susceptibility by disc diffusion method and molecular analysis of antibiotic resistance genes and certain virulence genes (PVL and fnBPA genes). RESULTS: Methicillin resistant S. aureus represented 178 (71%) of the isolated S. aureus and reduced susceptibility to vancomycin was detected by minimum inhibitory concentration in 39 (22%) isolates. It was found that there was a strong association between the MRSA strains and resistance to some antibiotics, devices association (p<0.001) and patient outcomes (p=0.003). There was a significant association between reduced vancomycin susceptibility (p=0.010), the presence of a central line catheter (p=0.000) and fnBPA gene (p<0.001) and mortality rate. CONCLUSIONS: The present study highlights that major S. aureus strains isolated from sepsis in pediatric patients were methicillin resistant with a substantial proportion of reduced susceptibility to vancomycin. Although none of the isolates had van genes responsible for vancomycin resistance, this finding warrants a considerable attention for study as it was a risk factor for mortality in those patients. The virulence genes fibronectin-binding protein and Panton-Valentine Leucocidin were not uncommon in S. aureus.