Production, Purification & Analysis of Enterotoxins from HA-MRSA Isolated from Clinical Samples

Author Name(s): K. Bharathi Bai, * Dr. S.S. Sudha
Author Email: Katewad.bharathi@gmail.com

Abstract

The problem of resistance is increasing for all types of bacterial isolates including Staphylococci. Methicillin resistant Staphylococcus aureus (MRSA) has emerged as an important hospital acquired and community acquired pathogen. Multi drug resistance among MRSA is a serious issue limiting the treatment options for the victims. MRSA strains that share the presence of staphylococcus cassette chromosome mec (SCC mec) in their genomes are frequently virulent and predominantly cause infections. Staphylococcal enterotoxins are a family of structurally related proteins that are produced by Staphylococcus aureus playing a major role in the pathogenicity. These microbial superantigens have profound effects on the immune system, which makes them useful tools for understanding its mechanism of action. The proposed study is mainly focused on collection of MRSA strains from clinical samples, isolation and Identification was done on the basis of microscopic examination and cultural characteristics, biochemical tests. The cultures were subjected for DNA extraction fallowed by AGE. All the samples were amplified targeting the gene Mec A and sent for sequencing. All the sequences were further analyzed and confirmed. The production and purification of Enterotoxins by MRSA was done and crude protein was collected. This was further purified by downstream processing. The sample was subjected for centrifugation followed by salt precipitation, Dialysis. The enterotoxins were produced form all the strains isolated and purified.

Introduction

Staphylococcus aureus is a critical human pathogen that can cause an assortment of illnesses, including skin and delicate tissue contaminations and orderly life undermine diseases. The proceeding with rise of methicillin resistant Staphylococcus aureus (MRSA), including hospital acquired (HA) – MRSA and community acquired (CA) – MRSA, is a noteworthy and expanding danger to general wellbeing. most of the nosocomial s. aureus infections are as a result of methicillin-resistant s. aureus (MRSA) lines and have turn out to be a broadly diagnosed motive of morbidity and mortality at some stage in the sector (ardic et al., 2006; pesavento et al., 2007; ho et al., 2008). In addition, MRSA lines proof against quinolones or multiresistant to other antibiotics have been emerging, leaving a restrained preference for their manipulate (mee-marquet et al., 2004; nejma et al., 2006; pesavento et al., 2007) The present study was the characterization of different staphylococcus aureus isolates collected from different clinical samples. Molecular level identification was done by DNA extraction fallowed AGE & then PCR based on 16s sequencing. This characterization was based on the ability of the isolates to produce and to express staphylococcal enterotoxins.

Conclusion

The above work concludes that all the bacteria isolated from the clinical samples was identified to be Staphylococcal sub species and are known fro the presence of Methicillin resistance gene. Also the work included the extraction and purification of enterotoxin that can have a therapeutic role in the further investigations.

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