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<h2>iGEM 2016</h2>
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<h4><strong>Project Description </strong></h4>
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<p><strong>Issue</strong></p>
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<p>In many developing countries, rapid urbanization leaves people with limited access to cooking facilities, resulting in a large dependence on reasonably priced and conveniently available street foods. These foods, however, pose a high risk of food poisoning due to microbial contamination. According to WHO statistics, food poisoning kills 420,000 people a year worldwide. One of the primary microbial contaminants is the Shiga toxin-producing E. coli O157:H7 (STEC). These particular pathogenic bacteria cause severe diseases in humans worldwide by secreting a toxin called Shiga-like toxin (SLT). Research shows that E. coli causes 73,000 illnesses in the United States every year. It is even estimated that STEC causes 2,801,000 acute illnesses worldwide annually, leading to 3890 cases of Hemolytic Uremic Syndrome, 270 cases of End-stage renal disease, and 230 deaths. Currently, there is no detection method for Shiga-like toxin outside of a lab setting, so consumers have no way of protecting themselves without strong legislation.</p>
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<p><strong>Project</strong></p>
 
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<p>Lack of action taken by governments and street food vendors in developing countries lead to the prevalence of street food-related illnesses, and call for the necessity of consumer awareness. Our project ultimately aims for a consumer-focused mechanism to detect Shiga-like toxins in foods.</p>
 
  
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<p>Shiga-like toxins are exotoxins, which consist of a toxic enzymatic A subunit and a cell-binding B subunit. The latter binds to a globotriaosylceramide (Gb3) receptor, expressed on the surface of the target cells. This interaction is responsible for the toxin's entry into the host cell. Through our device, Gb3 will be expressed in non-pathogenic E. coli. When the receptor is exposed to the food sample, the SLT subunit B, if present, will bind to Gb3. Crosslinking will occur to stabilize the interaction. Next, a recombinant subunit B will be fused with a reporter and then applied to detect whether the Gb3 binding sites are available. If the binding sites are vacant, the sub-unit B will bind to them and elicit a signal, indicating the safety of the food sample.</p>
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<p>In summary, we will be creating a portable device that would evaluate the safety of food by detecting a specific contaminant, Shiga-like toxin, for consumers who rely heavily on street food in developing countries in which there are weak food safety regulations.</p>
 
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<p><strong> Ideation Process</strong></p>
 
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<p>Before making our final decision, we explored an extremely diverse range of project topics such as blockage of oil pipes, controlling fruit ripening, detecting milk adulteration, biodegradable plastics, bioelectronics, preventing the coral reefs bleaching, bio-art, smoking, monitoring air quality, and more. After extensive research and intense discussion, we decided to move forward with the idea of detecting pathogenic SLT-producing E. coli in street foods. SLT is highly toxic due to its low LD50 and the A subunit’s protein synthesis inhibitory action, so it would have been too dangerous to work with given our lab environments. Keeping the validation process in mind, we decided to use only the SLT B subunit as our method of verification.</p>
 
  
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<li><a href="https://2016.igem.org/Team:NYU-AD/Description">Description</a></li>
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          <h1 id="project-abstract-heading">Project Abstract</h1>
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          <p id="abstract-para">In many developing countries, people depend on reasonably priced and conveniently available street food. However, lack of action taken by governments to regulate street food vendors has led to the prevalence of severe street food-related illnesses. One of the primary microbial contaminants in street food is <i>E. coli</i> O157:H7, which acts by secreting the Shiga-like toxin (SLT). Currently, there is no detection method for SLT outside of a lab setting, thus putting the consumers of foods at risk. Our project aims to develop a device that would be used by street vendors and restaurant owners to verify the safety of their products. Through our device, we exploit the binding of Gb3 to subunit B of the Shiga toxin, and compare the migration pattern of the bound Gb3-subunit B complex to a non bound subunit B. A shift in the migration pattern on a PAGE gel will occur when Gb3 is bound, indicating the presence of the toxin in the food sample. If no shift occurs in the SLT migration pattern, this implies the absence of the toxin within the sample, and reflects the safety status of the food.</p>
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            <h2 class="section-title"><span>Interested? Find out more about our...</span></h2>
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Latest revision as of 03:36, 3 December 2016

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Project Abstract

In many developing countries, people depend on reasonably priced and conveniently available street food. However, lack of action taken by governments to regulate street food vendors has led to the prevalence of severe street food-related illnesses. One of the primary microbial contaminants in street food is E. coli O157:H7, which acts by secreting the Shiga-like toxin (SLT). Currently, there is no detection method for SLT outside of a lab setting, thus putting the consumers of foods at risk. Our project aims to develop a device that would be used by street vendors and restaurant owners to verify the safety of their products. Through our device, we exploit the binding of Gb3 to subunit B of the Shiga toxin, and compare the migration pattern of the bound Gb3-subunit B complex to a non bound subunit B. A shift in the migration pattern on a PAGE gel will occur when Gb3 is bound, indicating the presence of the toxin in the food sample. If no shift occurs in the SLT migration pattern, this implies the absence of the toxin within the sample, and reflects the safety status of the food.



Interested? Find out more about our...


Project Description

Human Practices

Team




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