Difference between revisions of "Phage therapy"

Jump to navigation Jump to search
Line 104: Line 104:
== Questions and Specific Aims ==
== Questions and Specific Aims ==
:
:
:
:'''Pilot Project Aims'''
:
:'''Project Aims'''
<br />
<br />
1. Monitor the emergence of antibiotic resistant gonorrhea.<br />
'''Isolate N. gonorrhoeae from 10 patients'''. We are currently in negotiations with local clinics (ie. Berkeley Free Clinic, Magnet, City Clinic of San Francisco and St. James Infirmary) to enroll patients in our pilot study. Briefly, patients who are screened by these clinics and test positive for oropharyngeal N. gonorrhoeae will be provided with information on our study. Within the informational packet, patients will find their assigned Patient ID and use this number to schedule an in-house visit. The Patient ID allows the patient to remain anonymous during our screening process. During their primary visit, patients will provide their consent, sign liability waivers, fill out our surveillance survey, and provide us with an oropharyngeal specimen. Patients can then use their Patient ID to track the results from their specimen using our anonymous web tracking system.
:a. Compose and deploy a mobile screening unit at various locations throughout Oakland and San Francisco.
 
:b. Compile data from the mobile screening program and integrate it with data from other public health organizations.
Primary gonococcal cultures will be grown on Modified Thayer-Martin (MTM) medium containing chocolate agar (5% SRBC), Vancomycin, Colistin, Nystatin and SXT. This medium allows selective growth of Neisseria species. Primary cultures will be propagated in tryptic soy broth (TSB) according to the standards set forth by the Centers for Disease Control (REF).  
<br />
 
2. Develop a community laboratory space in Oakland, CA that has the equipment we need in which we can conduct our research.
Upon primary culture and propagation, N. gonorrhoeae samples will be confirmed using nucleic acid testing. A panel of polymerase chain reactions will be performed on DNA isolated from bacterial cultures. These panels will be performed to (1) identify genes responsible for 1st-, 2nd- and 3rd-generation antibiotic resistance, (2) differentiate N. gonorrhoeae from other Neisseria species. The differentiation PCR will be confirmed using CTA sugar growth testing as described by the CDC (REF). Briefly, isolates will be assessed for their ability to grow in Glucose, Maltose, Lactose and Sucrose. N. gonorrhoeae is able to only grow in glucose.
:a. Procure capital equipment and reagents needed for this project.  
 
:b. Procure reagents and equipment to begin molecular microbiological research.
'''Isolate ARNG-derived bacteriophage from 10 patients'''.  
<br />
 
3. Isolate, identify and characterize the first known bacteriophages from '''N. gonorrhoeae.'''
'''Characterize ARNG-derived bacteriophage from 10 patients'''.
:a. Harvest samples from our mobile screening program.
 
:b. Perform Genomic and Proteomic Analyses on Bacteriophage samples.
'''Conduct proof-of-concept lysis experiments'''.
<br />
4.Develop methods by which bacteriophages can be cultured and used as prophylaxis and therapy for gonoccocal infections.
:a. Test and optimize growth conditions for strains of antibiotic resistant gonorrhea from which bacteriophage can be isolated.
:b. Harvest phage and perform proof-of-concept, in vitro experiments in which gonorrhea is lysed with preparations of bacteriophage.
<br />
5. Provide educational outreach opportunties.
:a. Provide information on prevention and treatment of gonorrhea.
:b. Provide in-the-lab training and experience for those with the desire to pursue citizen science.


== Anticipated Challenges to Our Approach ==
== Anticipated Challenges to Our Approach ==