Evaluation of icaA and icaD genes involved in biofilm formation in Staphylococcus aureus isolates from clinical sources using Reverse Transcriptase PCR

Document Type : Original Articles

Authors

1 Biotechnology Research Center, Karaj Branch, Islamic Azad University, Karaj, Iran

2 Molecular Biology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran

3 Department of Microbiology, Faculty of Medicine, Guilan University of Medical Sciences, Rasht, Iran

10.22092/ari.2024.364437.2979

Abstract

Staphylococcus aureus is known for its ability to create biofilm, which helps bacteria attach to various surfaces and poses a challenge for treatment. Biofilm formation relies on the icaABCD operon, with the icaA and icaD genes having the key role in this complex process. The aim of the study is to investigate the role of these genes in the biofilm formation of S. aureus isolates sourced from clinical settings. We collected 100 S. aureus isolates from clinical sources, then extracted DNA and RNA using a commercial kit from Kiagen Co. To transcribe the RNA samples into cDNA, we used a commercial kit from Kiagen Co. We measured the ability to produce phenotypic and molecular biofilm formation using the microtiter plate method and PCR, respectively. We were determining the expression of icaA and icaD genes via RT-PCR (Reverse transcription polymerase chain reaction). Ninety-five percent (95%) of the isolates were able to produce biofilm, with 16 (16%) producing weak, 64 (64%) producing medium, and 15 (15%) producing strong biofilms. Additionally, 72 (72%) of the isolates had the icaA gene, while 58 (58%) carried the icaD gene. Out of these isolates, 70 (97.2%) isolates expressed the icaA gene, and 53 (73.6%) isolates expressed the icaD gene. Four isolates (5.5%) that had the icaA gene but lacked the icaD gene did not form biofilm. One strain did not express either of the genes. The presence of either the icaA or icaD gene is crucial for the development of biofilm. However, further investigation is needed to fully understand the formation of biofilm.

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Archives of Razi Institute

Articles in Press, Accepted Manuscript
Available Online from 06 May 2024

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