" Simplex, Duplex and Multiplex PCR for detection up to Six Enterococcus virulence genes in Staphylococcus spp. isolated from burn patients"

The high incidence of Gram-positive cocci particularly Staphylococcus aureus poses a severe threat in burn patients, causing a wide variety of diseases ranging from minor skin infection to life-threatening sepsis cases, leading to infection treatment difficulties. This study aimed to investigate genotypic approach to detect six virulence genes in Staphylococcus spp. may passed from Enterococcus spp. isolated from the same niches of burn patients. The study analyzed 200 clinical samples (wound, urine, stool, and blood) from 75 burn patients. The chosen six virulence genes including: asa1, gelE, cylA, esp, hyl, and efa, were detected using simplex, duplex and multiplex PCR alongside phenotypic in Staphylococcus spp., in addition to antibiotics resistance genes vanA and aph(3’). Initial identification used selective media and biochemical tests, followed by 16S rRNA sequencing, of these samples, 38 (19%) were confirmed as Staphylococcus species. The distribution included S. epidermidis (31.5%), S. aureus (28.5%), S. haemolyticus (23.6%), S. hominis (13%), and S. saprophyticus (2.6%). Notably, all 38 isolates grew on HiChrome media typically specific for Enterococcus faecium. The asa1 gene was the most prevalent, it was recorded in up to 100% of isolates with significant differences p<0.0001. Other genes like gelE and hyl also showed high frequency, while efa, esp, and cylA levels fluctuated across different PCR methods. A key finding the coexistence of these two bacterial species in the same niche raises concerns about the potential horizontal transfer of virulence-associated traits between them, highlighting the importance of rapid and accurate diagnosis to minimize this potential risk.