DETERMINATION OF BIOFILM FORMATION GENES USING PCR TECHNIQUE FOR STAPH. SPP. ISOLATIONS FROM WOUND AND BURN INFECTIONS IN BAQUBA CITY

Abstract

The bacteria Staphylococcus aureus has been discovered to be a major source of community and hospital-acquired infections. The production of ica-dependent biofilms is critical in the persistence of infections in hospitalized patients. Between November 2017 &April 2018, the current study was conducted at Teaching Baquba Hospital's Bacteriology Laboratory in Baquba City and the laboratory of microbiology and polymerase chain reaction (PCR )unit in the Biology Department / College of Science/ Diyala University (2018). Materials and methods: We obtained 13(17.3%) Staph.aureus isolates from 100 clinical specimens (burns, wounds, urine, and blood) after identified them. Following by employed Congo Red Agar(CRA) and tissue culture plate method (TCP)to detect Biofilm development in isolates, as well as a PCR assay and particular primers to determine the presence of the icaA &icaD genes. The results showed ica A/D were found in 69 % (9/13) of cases, icaA gene is present at 7 (53.8%) and the icaD gene at 2(15 .3%) in Staph.aureus isolates. CRA method found biofilm generation in 6 (46%) of thirteen Staph. aureus isolates, while TCP detected biofilm creation in 10 (76%) isolates. When phenotypic approaches compared to the detection of the icaA and icaD genes, only 5 (71%) of the icaA genes were found to be positive by TCP, while only 2 (1% ) of the icaD genes were found to be positive by TCP. In short: The findings show the significance of S. aureus' virulence factors in clinical samples for the icaA and icaD genes and the phenotypic biofilm formation variety. The creation of in vitro slime using the CRA approach is not necessarily consistent even when the icaA and icaD genes exist. Although certain isolates lack the genes icaA & icaD, the ability to generate biofilms highlights the importance of the further gene research, and the absence of the icaA and icaD genes, the capability from certain isolates to create biopolymes emphasises the need for continuous genetic study into icas caused by variations in the number of genes associated with biofilms. When comparing phenotypic techniques, TCP is still the best tool for the screening of biofilms. The aim of this research though is that the biofilm forming potential should be actually linked to the presence of icaA and icaD genes in S. aureus isolates