• Edwina Jospiene M
• Dr.S. Subramaniam
• Dr. Shyama Subramaniam

Staphylococcus aureus is the major contagious nosocomial pathogen and has no or limited rapid diagnostic tools. Strain variation and little knowledge of common immunogenic antigens are among the major constraints to developing effective diagnostic tools. This study aimed to compare four novel Staphylococcus aureus protein extraction methods and evaluate the immune reactivity of extracted proteins. After extraction, bacterial cell integrity was examined by Gram staining and light microscopy to determine whether the extraction methods caused damage to cells that may release non-surface proteins. The extracted proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and evaluated for immune reactivity using western blot. Results show that all four extraction methods provided multiple protein bands on SDS-PAGE. Western blot results showed several immunoreactive surface proteins, in which some proteins strongly (well-resolved, thick, dark, and intense band) reacted to the tested. The confirmation of the species was done by MALDI TOF MS as MRSA. Hence the proteins isolated in this study are specific to the MRSA species of S.Aureus. All four methods are valid for extracting surface proteins, the hexadecane method is the most feasible. All four methods are easy, cheap, and have minor effects on the bacterial cell wall. Strongly immune-reactive proteins will serve as potential candidates as immunohistochemistry markers.

Quick, Bacterial protein extraction, Staphylococcus aureus, SDS, Western blot

"Rapid Protein Extraction Method; A Method Comparison Study", International Journal of Emerging Technologies and Innovative Research (www.jetir.org), ISSN:2349-5162, Vol.10, Issue 8, page no.b480-b488, August-2023, Available :http://www.jetir.org/papers/JETIR2308159.pdf

"Rapid Protein Extraction Method; A Method Comparison Study", International Journal of Emerging Technologies and Innovative Research (www.jetir.org | UGC and issn Approved), ISSN:2349-5162, Vol.10, Issue 8, page no. ppb480-b488, August-2023, Available at : http://www.jetir.org/papers/JETIR2308159.pdf