Team:LMU-Munich/Application/Diagnosis Treatment

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= Diagnostics & Treatment =
= Diagnostics & Treatment =
== Treatment ==
== Treatment ==
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[[File:LMU2014 Treatment Bakillus Illustrationsgrafik 01.jpg|960px]]
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[[File:LMU2014 Treatment Bakillus Illustrationsgrafik 01.jpg|thumb|middle|500px|Figure 1: Draft of the BaKillus Nasal Spray]]
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The human nasal mucosa is the main ecological niche of ''Staphylococcus aureus''  residing in human beings.  ''S. aureus'' adheres to the mucosa and the epithelia of the anterior nares. The goal of BaKillus is to decolonize and kill ''S. aureus'' within the interior nasal areas. Other methods of decolonizing ''S. aureus'' have already shown to reduce the risk of infections with ''S. aureus''.
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The human nasal mucosa is the main ecological niche of ''Staphylococcus aureus''  residing in human beings.  ''S. aureus'' adheres to the mucosa and the epithelia of the anterior nares[http://dx.doi.org/10.1016/S0966-842X(01)02254-5]. The goal of BaKillus is to decolonize and kill ''S. aureus'' within the interior nasal areas. Other methods of decolonizing ''S. aureus'' have already shown to reduce the risk of infections with ''S. aureus'' subsequently to open surgery[http://dx.doi.org/10.1056/NEJM200101043440102][http://dx.doi.org/NEJMoa0808939]. Nonetheless those current preventive measures include efforts towards screening patients that will receive surgery in order to avoid overusage of antibiotics.
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BaKillus resembles an alternative possibility as it will only produce antibiotics in response to the actual presence of ''Staphylococcus aureus''. It is therefore possible to apply BaKillus as an preventive application without need for screening and unneccessary usage of antibiotics at the same time.
To maintain a cheap and easy-to-use application we decided to create a specialized nasal spray. The general functionality is the same as for any common nasal spray for medical applications, so no pre-experience is needed to use our BaKillus Spray. The characteristic of ''Bacillus subtilis'' to build heat resistant spores, that can be stored dryly gives the spray a very high life period as long as the spores are not activated (learn more about [https://2014.igem.org/Team:LMU-Munich/Project/B_subtilis ''Bacillus subtilis'']).
To maintain a cheap and easy-to-use application we decided to create a specialized nasal spray. The general functionality is the same as for any common nasal spray for medical applications, so no pre-experience is needed to use our BaKillus Spray. The characteristic of ''Bacillus subtilis'' to build heat resistant spores, that can be stored dryly gives the spray a very high life period as long as the spores are not activated (learn more about [https://2014.igem.org/Team:LMU-Munich/Project/B_subtilis ''Bacillus subtilis'']).
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On the first pumping, the lamination will be perforated by being pushed onto the cutting edge ring. The spores will become free and mix into the second chamber below. Inside this chamber, sterile defined media as well as an oxygen-enriched atmosphere will be presented to the spores. Application of the activated BaKillus spray to the nostrils will be identical to common nasal sprays.
On the first pumping, the lamination will be perforated by being pushed onto the cutting edge ring. The spores will become free and mix into the second chamber below. Inside this chamber, sterile defined media as well as an oxygen-enriched atmosphere will be presented to the spores. Application of the activated BaKillus spray to the nostrils will be identical to common nasal sprays.
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Usage of the BaKillus Spray is described below:
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<br>Usage of the BaKillus Spray is described below:
* Start spore activation by pushing down the pumping mechanism once to break the lamination seal.
* Start spore activation by pushing down the pumping mechanism once to break the lamination seal.
* Mix the spores and the medium by shaking vigorously.
* Mix the spores and the medium by shaking vigorously.
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For the future we emphasize a rent-and-return system for the nasal spray itself in combination with a more dispersed distributional network. For more information about future application scenarios please visit our (LINK: Collaboration with the Rathenau Institute) page.
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For the future we emphasize a rent-and-return system for the nasal spray itself in combination with a more dispersed distributional network. For more information about future application scenarios please visit our [https://2014.igem.org/Team:LMU-Munich/Rathenau Collaboration with the Rathenau Institute] page.
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For a second application, using a similar principle we invite you to visit our collaboration partner's Wiki from Groningen about the Lacto Aid System. (LINK TO COLLABORATION PAGE)
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For a second application, using a similar principle we invite you to visit our [https://2014.igem.org/Team:LMU-Munich/Team/Collaborations collaboration] partner's Wiki from Groningen about the Lacto Aid System.
== Diagnosis ==
== Diagnosis ==
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<b>Problem:</b>
<b>Problem:</b>
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Current tests to identify S. aureus are PCR-based to detect specific genes. Theoretically, a PCR result could be obtained within three hours, whereas it would take around seven hours in practice. However, PCR-based methods are error-prone. Therefore, verification by a molecular biological test is recommended. Current biological tests (e.g. coagulase) require a time-consuming (18-24h) culturing step. Based on the test results, rapid and strict hygiene rules, e.g. separation from non-infected persons, seem to be very effective to lower the spread of health-care associated MRSA.  
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Current tests to identify S. aureus are PCR-based detections of pathogene-specific genes. Theoretically, a PCR result could be obtained within three hours, whereas it takes around seven hours in practice. However, PCR-based methods are error-prone. Therefore, verification by a molecular biological test is recommended. Current biological tests (e.g. coagulase) require a time-consuming (18-24h) culturing step. Based on the test results, rapid and strict hygiene rules, e.g. separation from non-infected persons, seem to be very effective to lower the spread of health-care associated MRSA.  
Here, we suggest an alternative strategy that can potentially facilitate the current diagnosis procedure.
Here, we suggest an alternative strategy that can potentially facilitate the current diagnosis procedure.
<b>The BaKillus S. aureus diagnosis tool:</b>
<b>The BaKillus S. aureus diagnosis tool:</b>
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We are planning to develop a ready-to use diagnosis tool for point-of-care testing by the professional staff in hospitals, pharmacies or medical offices (see Fig. 2). A smear of a patient into a BaKillus-reporter culture could be enough to verify the presence of S. aureus within 0.5-2 hours (just limited by the time for BaKillus spore germination and reporter gene expression). This culture-independent detection could be precautionary used by patients of certain risk groups. The fast result would enable health-care institutions to start directly a subsequent treatments/hygiene measure just after arrival of the patient and thus contact between infected and non-infected can be lowered.
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We are planning to develop a ready-to-use diagnosis tool for point-of-care testing by the professional staff in hospitals, pharmacies or medical offices (see Fig. 2). A smear of a patient into a BaKillus-reporter culture could be enough to verify the presence of ''S. aureus'' within 0.5-2 hours (just limited by the time for BaKillus spore germination and reporter gene expression). This culture-independent detection could be precautionary used by patients of certain risk groups. The fast result would enable health-care institutions to directly start subsequent treatments/hygiene measures just after arrival of the patient and thus contact between infected and non-infected patients and therefore the spreading of multi-resistant pathogens could be lowered.
[[File:LMU14_Application_Rathenau_Diagnostic_Device.png|thumb|700px|center|
[[File:LMU14_Application_Rathenau_Diagnostic_Device.png|thumb|700px|center|
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Figure 2: Basic concept of the BaKillus diagnostic tool. BaKillus spores are stored separately in a self-contained device. About one hour before use, spores and medium are mixed and incubated for the germination of the spores. The sample of a patient (e.g. a smear) can be applied into the culture and after about 30 minutes the result can be analysed with a simple camera-computer-station.
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Figure 2: Basic concept of the BaKillus diagnostic tool. BaKillus spores are stored separately in a self-contained device. About one hour before use, spores and medium are mixed and incubated for the germination of the spores. The sample of a patient (e.g. a smear) can be applied into the culture and after about 30 minutes the result can be analysed (e.g. pigment production or luminescence as a reaction to pathogens)
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<i>Table 1: Discussion of the BaKillus diagnostic tool.</i>
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{|
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{| class="wikitable"
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|+ Table 1: Discussion of the BaKillus diagnostic tool.
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|-
|-
!Strengths
!Strengths
!Weaknesses
!Weaknesses
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|-
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|Detection & Identification of AIP-dependent pathogens  
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|Detection & Identification of AIP-dependent pathogens discriminating them from the opportunistic non-pathogenic state
|Application is limited to targets that exhibit QS-dependent pathogenicity.  
|Application is limited to targets that exhibit QS-dependent pathogenicity.  
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|-
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| Pigments (https://2012.igem.org/Team:Groningen/pigmentproduction) would be ideal to use as bio-reporters (the output of the detection device). The change of colour can be observed by eye and thus the product needs no high-end technology or electricity =>
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|[https://2012.igem.org/Team:Groningen/pigmentproduction Pigments] would be ideal to use as bio-reporters (the output of the detection device). The change of colour can be observed by eye and thus the product needs no high-end technology or electricity. Therefore highly portable and independent device for usage in abroad and crisis regions
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Highly portable and independent device for usage in outlands and crisis regions
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|Initial assessment of the sensitivity of the test and tests for reliability of the tool required  
|Initial assessment of the sensitivity of the test and tests for reliability of the tool required  
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Latest revision as of 13:52, 10 December 2014

 

Diagnostics & Treatment

Treatment

Figure 1: Draft of the BaKillus Nasal Spray

The human nasal mucosa is the main ecological niche of Staphylococcus aureus residing in human beings. S. aureus adheres to the mucosa and the epithelia of the anterior nares[1]. The goal of BaKillus is to decolonize and kill S. aureus within the interior nasal areas. Other methods of decolonizing S. aureus have already shown to reduce the risk of infections with S. aureus subsequently to open surgery[2][3]. Nonetheless those current preventive measures include efforts towards screening patients that will receive surgery in order to avoid overusage of antibiotics. BaKillus resembles an alternative possibility as it will only produce antibiotics in response to the actual presence of Staphylococcus aureus. It is therefore possible to apply BaKillus as an preventive application without need for screening and unneccessary usage of antibiotics at the same time.

To maintain a cheap and easy-to-use application we decided to create a specialized nasal spray. The general functionality is the same as for any common nasal spray for medical applications, so no pre-experience is needed to use our BaKillus Spray. The characteristic of Bacillus subtilis to build heat resistant spores, that can be stored dryly gives the spray a very high life period as long as the spores are not activated (learn more about Bacillus subtilis). To make sure the BaKillus spores do not get activated too early we developed an additional chamber associated to the pump mechanism. This chamber in the shape of an open ring (Fig. 1) will be sealed by a two component lamination. One layer of plastics will tighten the chamber while a cellulose layer will add stability and make the lamination perforable. BaKillus spores will be enclosed within the chamber. In the assembled product On the first pumping, the lamination will be perforated by being pushed onto the cutting edge ring. The spores will become free and mix into the second chamber below. Inside this chamber, sterile defined media as well as an oxygen-enriched atmosphere will be presented to the spores. Application of the activated BaKillus spray to the nostrils will be identical to common nasal sprays.


Usage of the BaKillus Spray is described below:

  • Start spore activation by pushing down the pumping mechanism once to break the lamination seal.
  • Mix the spores and the medium by shaking vigorously.
  • Incubate at 37 °C for 2 hours to activate the spores to the BaKillus cells.
  • Apply BaKillus Spray to each nostril every 2 hours. Reactivated Spores can be used for 20 hours before they will yield useless.
  • Recycle the Nasal Spray.


For the future we emphasize a rent-and-return system for the nasal spray itself in combination with a more dispersed distributional network. For more information about future application scenarios please visit our Collaboration with the Rathenau Institute page.

For a second application, using a similar principle we invite you to visit our collaboration partner's Wiki from Groningen about the Lacto Aid System.

Diagnosis

Pathogen-identification diagnostic tool:

Problem:

Current tests to identify S. aureus are PCR-based detections of pathogene-specific genes. Theoretically, a PCR result could be obtained within three hours, whereas it takes around seven hours in practice. However, PCR-based methods are error-prone. Therefore, verification by a molecular biological test is recommended. Current biological tests (e.g. coagulase) require a time-consuming (18-24h) culturing step. Based on the test results, rapid and strict hygiene rules, e.g. separation from non-infected persons, seem to be very effective to lower the spread of health-care associated MRSA.

Here, we suggest an alternative strategy that can potentially facilitate the current diagnosis procedure.

The BaKillus S. aureus diagnosis tool:

We are planning to develop a ready-to-use diagnosis tool for point-of-care testing by the professional staff in hospitals, pharmacies or medical offices (see Fig. 2). A smear of a patient into a BaKillus-reporter culture could be enough to verify the presence of S. aureus within 0.5-2 hours (just limited by the time for BaKillus spore germination and reporter gene expression). This culture-independent detection could be precautionary used by patients of certain risk groups. The fast result would enable health-care institutions to directly start subsequent treatments/hygiene measures just after arrival of the patient and thus contact between infected and non-infected patients and therefore the spreading of multi-resistant pathogens could be lowered.

Figure 2: Basic concept of the BaKillus diagnostic tool. BaKillus spores are stored separately in a self-contained device. About one hour before use, spores and medium are mixed and incubated for the germination of the spores. The sample of a patient (e.g. a smear) can be applied into the culture and after about 30 minutes the result can be analysed (e.g. pigment production or luminescence as a reaction to pathogens)


Table 1: Discussion of the BaKillus diagnostic tool.

Strengths Weaknesses
Detection & Identification of AIP-dependent pathogens discriminating them from the opportunistic non-pathogenic state Application is limited to targets that exhibit QS-dependent pathogenicity.
Pigments would be ideal to use as bio-reporters (the output of the detection device). The change of colour can be observed by eye and thus the product needs no high-end technology or electricity. Therefore highly portable and independent device for usage in abroad and crisis regions Initial assessment of the sensitivity of the test and tests for reliability of the tool required
Self-contained device prevents release of GMOs and thus the product only requires a proposal for release (personal communication Dr. Ulrich Ehlers, Commissioner for GMO-release in Germany, BVL and Dr. Hans Schrubar Commissioner for Bio-Safety in Bavaria) German laws are unspecific about bacterial GMOs. Success and risks are hard to calculate

Hi there!

Welcome to our Wiki! I'm BaKillus, the pathogen-hunting microbe, and I'll guide you on this tour through our project. If you want to learn more about a specific step, you can simply close the tour and come back to it anytime you like. So let's start!

What's the problem?

First of all, what am I doing here? The problem is, pathogenic bacteria all around the world are becoming more and more resistant against antimicrobial drugs. One major reason for the trend is the inappropriate use of drugs. With my BaKillus super powers, I want to reduce this misuse and thus do my part to save global health.

Sensing of pathogens

To combat the pathogenic bacteria, I simply eavesdrop on their communication. Bacteria talk with each other via quorum sensing systems, which I use to detect them and trigger my responses.

Adhesion

The more specific and effective I can use my powers, the lower the danger is of provoking new resistance development. So I catch pathogens whenever I get hold of them and stick to them until my work is done.

Killing

Talking about my work - killing pathogens is finally what I am made for. In response to quorum sensing molecules of the pathogens, I export a range of antimicrobial substances leading to dissipation of biofilms and the killing of the targeted bacteria.

Suicide switch

When the job is done and all the bad guys are finished, you don't need a super hero anymore. So after fulfilling my work I say goodbye to the world by activating my suicide switch.

Application

Of course I'm not only a fictional hero, but a very real one. In two different prototypes, I could be used for diagnosis or treatment of pathogen-caused diseases. However, there is still a whole lot of regulational and economical questions that have to be answered before.

See you!

So now you know my short story - and it is time for me to return to my fight for a safer world. Feel free to take a closer look on my super powers, the process of my development or the plans for a medical application.