Difference between revisions of "Team:UiOslo Norway/Description"

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Our goal is to make a diagnostic test associated with an app, which is available for doctors both in the field and in the medical offices. We’re in addition making a scheme of an screening test that fits the mention criteria.

Diagnostic test: We will make a diagnostic test that will detect ESBL producing bacteria in urinary tract infections. The already established Penta Well test by Mura et. al. (2015), changes color from yellow to red when this resistant bacterium is present. The color change is due to the cleavage of the betalactam-ring in Nitrocefin. The test can also discriminate between several classes of betalactamases, and we intend to base our idea on this.

Smartphone app: The diagnostic test will be associated with a smartphone app – phonelab. The camera on your smartphone will be connected to the app which by help of a binary code will detect any colorchange. Color will be cross-referenced with a database which will give the doctor a list of any resistant bacteria found and what antibiotics not to give.

Screening test: We want to use a simplified CRISPR/Cas9 system to link base-pairing to a detectable signal. The system will be comprised of a gRNA towards the dangerous resistance genes and a modified Cas9 protein. This will enable us to screen for the dangerous resistance genes, thus preventing antibiotics to end up in the wrong place even before the resistance gene products are already present. Conceptually; the gRNA is linked to a null-nuclease Cas9 that doesn´t cleave the target genes. Linked to the Cas9 is a detectable partner, potentially another protein. When gRNA binds to its target, Cas9 functions as a link between the gene and its partner. The bottleneck right now is to think of a detection method that will only detect the Cas9-construct when bound to the genes while being fast and easy enough to be relevant for our idea.

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-This year’s iGEM team from UiO will address one of the increasing world problems, antibiotic resistance. For many years humans have evolved together with bacteria in an evolutionary race, for each new antibiotic produced, a simple bacteria will eventually be able to resist the medicine, survive the treatment and continue to grow. Today we have come to a place where there is little or no new antibiotics produced and the resistance of the ones that are in use is increasing.  Without functional antibiotics modern medicine will fall back many centuries. Without antibiotics doctors won’t be able to perform basic surgeries, keep diabetes patients healthy, and treat pre – term babies from infections. It is postulated that antibiotic resistance may become a greater threat to our health than cancer, and by 2050 10 million people is estimated to die due to infections caused by antibiotic resistant bacteria. </p>
+Our goal is to make a diagnostic test associated with an app, which is available for doctors both in the field and in the medical offices. We’re in addition making a scheme of an screening test that fits the mention criteria.
+</p>
 <p>
-A part that contributes to the increased antibiotic resistance is the unnecessary use of antibiotics. It may be prescribed when patients come in with virus infections, or infections that will go over with time or it may not even be prescribed at all. In developing countries it’s possible to buy antibiotics with no diagnosis of disease or infections. In addition, the factories that produce these drugs do not have an appropriate waste policy and antibiotics are released into the nearby area, for example in rivers where people collect drinking water.  (Norwegian TV- program (brennpunkt)). All these things will provide bacteria a selective pressure which will select for the survival of the ones that gains antibiotic resistance and they will continue to spread. </p>
+Diagnostic test:  We will make a diagnostic test that will detect ESBL producing bacteria in urinary tract infections. The already established Penta Well test by Mura et. al. (2015), changes color from yellow to red when this resistant bacterium is present. The color change is due to the cleavage of the betalactam-ring in Nitrocefin. The test can also discriminate between several classes of betalactamases, and we intend to base our idea on this.
+</p>
-<p> With this project we want to make people aware of this increasing problem, by taking advantage of social media, give lectures and attend several science fairs. </p>
-<p>How is it that a simple organism such as bacteria can pose such a big threat to mankind? It’s important not to underestimate these prokaryotic organisms. They were among the first to habitat this earth and have evolved to survive in almost every corner of the world. They have a unique ability to take up DNA either from other bacteria or free DNA, in this way they can obtain new genes that may give them some evolutionary advantage, and by natural selection these will survive. By obtaining these new genes some will give bacteria a clever mechanism to escape the actions of antibiotics. We have focused our project to address one of these mechanisms, the production of an enzyme that cleaves the antibiotics. This is one example of how bacteria can become resistance to the antibiotic Penicillin. This drug was the first antibiotic to be discovered and is still widely used today. Penicillin consist of a beta – lactam ring, this ring will bind to an enzyme (DD – transpeptidase) that is in charge of renewing the bacterial cell wall. Without this enzyme there will be no new formations of peptidoglycans for the cell wall and it will eventually rupture and the bacteria will die.</p>
+<p>
+Smartphone app: The diagnostic test will be associated with a smartphone app – phonelab. The camera on your smartphone will be connected to the app which by help of a binary code will detect any colorchange. Color will be cross-referenced with a database which will give the doctor a list of any resistant bacteria found and what antibiotics not to give.
-<p>When a bacterium is resistant to Penicillin it produces an enzyme called beta - lactamase. This enzyme will hydrolyse the beta lactam ring, making it unable to bind to the cell wall producing enzyme, thus the Penicillin will lose its destructive activity.</p>
-<p>In addition to inform the people of this imposing threat we will make a simple detection method for Penicillin resistance that can be used in every doctor’s office before any antibiotics are prescribed to the patient. We have focused on urinary tract infections which are very common (need numbers here) caused by ESBL producing E.Coli or Klebsiella spp.  We will accomplish this by making an assay that will detect the beta lactamase activity. (Or the gene with CRISPR/CAS)
 </p>
 <p>
-To detect this enzyme activity we will generate a device that can measure pH/colour change when the beta lactam ring is hydrolysed. If pH:  When this ring is hydrolysed it will release a proton which will acidify the solution.  If colour change: The activity can be detected by measuring the change in absorbance when adding a Penicillin derivative called Nitrocefin, this compound also has a beta lactam ring and appear  yellow. This compound usually has an absorption peak at 386nm, but when it’s hydrolysed it’ll change colour to red and wavelength can be measured at 482nm. (Or if contact with serum 510nm),(source: “Novel method for detection of B-lactamases by using a chromogenic cephalosporin substrate”,  O’ Callaghan, 1971).</p>
+Screening test: We want to use a simplified CRISPR/Cas9 system to link base-pairing to a detectable signal. The system will be comprised of a gRNA towards the dangerous resistance genes and a modified Cas9 protein. This will enable us to screen for the dangerous resistance genes, thus preventing antibiotics to end up in the wrong place even before the resistance gene products are already present. Conceptually; the gRNA is linked to a null-nuclease Cas9 that doesn´t cleave the target genes. Linked to the Cas9 is a detectable partner, potentially another protein. When gRNA binds to its target, Cas9 functions as a link between the gene and its partner. The bottleneck right now is to think of a detection method that will only detect the Cas9-construct when bound to the genes while being fast and easy enough to be relevant for our idea.
+</p>
 </div>