Development of Multiplex PCR for Rapid Detection of Toxigenic Strains of Staphylococcus aureus from Food Samples

Author Name: *D. Gayathri ,M. Shiva Prakash
Author Email: chilly79in@yahoo.co.in

Abstract

This study developed a multiplex PCR method for the screening and detection of five toxigenic strains of Staphylococcus aureus. The multiplex PCR procedure, which uses five pairs of primers produced specific amplicon of expected sized of SEA, SEB, SEC, SED and SEE genes which are responsible for producing Enterotoxins. These are the target genes for m-PCR detection. Overall results for the present study indicates that m-PCR is a potential technique for the rapid detection of all the five toxin producing strains of Staphylococcus aureus, which is one of the pathogenic food borne bacteria for routine monitoring and risk assessment of foods.

Keywords

Staphylococcus aureus, Food Borne pathogen, Multiplex PCR

Introduction

Staphylococcus aureus is a bacterial pathogen considered a principle etiological agent of food poisoning. They are the second most common pathogen associated with outbreaks of food poisoning [1].Staphylococcus aureus, a Gram positive bacterium is one of the most common clinical and food borne pathogens worldwide. Staphylococcus aureus produces many important virulence factors including Staphylococcus enterotoxins SEs which are the main causes of diarrhea, vomiting and other symptoms associated with Staphylococcus aureus infections [2]. There are several distinct serological enterotoxins(SE) such as SEA, SEB, SEC, SED, SEE, SEG, SEH, SEI, SES, SET. Out of all the Staphylococcus aureus enterotoxin SEA, SEB, SEC, SED, SEE showed emetic activity [3].The biological effects of all these enterotoxins include pyrogenicity, enhancement of lethal endotoxin shock and inflammatory cytotoxins. Staphylococcus aureus causes enterocolitis [4] Microbial and immulogical techniques, presently in use are time consuming and laborious. In addition the toxin escape during sterile filtration are heat resistant and therefore this necessitates the development of more sensitive technique for identifying the toxigenic Staphylococcus aureus. Therefore early identification
by this method will help in minimizing the infections there by preventing diarrheal diseases. This technique will also be useful in the food testing laboratories and medical diagnostic laboratories by health centers and at field levels.

Conclusion

Staphylococcal infection is currently widespread throughout the world and has prompt interest and concern for the rapid detection of toxigenic strains of Staphylococcus aureus. Staphylococcus aureus is a major human pathogen and causes a variety of nosocomial and community acquired infections. Molecular based methods have well grounded potential to overcome insufficiencies of identification procedures associated with the results based on biochemical characteristics. Developed PCR techniques on further investigations focus on the application to large scale of clinical samples so that this technique can be used in detecting toxigenic strain of Staphylococcus aureus for a public safety concern in foods.

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