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Difference between revisions of "Phage therapy"
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:'''Craig Rouskey''', MSc in Molecular Biology, Biochemistry, Microbiology and Immunology | :'''Craig Rouskey''', MSc in Molecular Biology, Biochemistry, Microbiology and Immunology | ||
:'''Ami Knop Ullrich''', MSc in Medical Microbiology and Immunology | :'''Ami Knop Ullrich''', MSc in Medical Microbiology and Immunology | ||
== Introduction == | == Introduction == | ||
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== Questions and Specific Aims == | == Questions and Specific Aims == | ||
[[Experiments]] | |||
:'''Pilot Project Aims''' | :'''Pilot Project Aims''' | ||
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:''Confirmation of N. gonorrhoeae''. 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 positive for glucose metabolism, but not maltose or lactose metablism. | :''Confirmation of N. gonorrhoeae''. 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 positive for glucose metabolism, but not maltose or lactose metablism. | ||
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:'''Isolate ARNG-derived bacteriophage from 10 patients'''. | :'''Isolate ARNG-derived bacteriophage from 10 patients'''. | ||
:''Bacteriophage Isolation''. Bacteriophage will be isolated from 10 clinical samples as previously described (REF). Briefly, isolated colonies will be selected and cultured in 5 mL TPY broth (trypticase, phytone, and yeast extract) and cultued for 24 hours at 37oC in 5% CO2. | :''Bacteriophage Isolation''. Bacteriophage will be isolated from 10 clinical samples as previously described (REF). Briefly, isolated colonies will be selected and cultured in 5 mL TPY broth (trypticase, phytone, and yeast extract) and cultued for 24 hours at 37oC in 5% CO2. | ||
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#Addressing previous citizen science failures and conducting research with a greater adherence to quality. | #Addressing previous citizen science failures and conducting research with a greater adherence to quality. | ||
#Preventing Lysogeny. | #Preventing Lysogeny. | ||
===Protocol Documentation=== | ===Protocol Documentation=== | ||
http://aegia.nu/130721_prephage-protocols.txt | http://aegia.nu/folder/getitproject/130721_prephage-protocols.txt | ||
== Laboratory Needs == | == Laboratory Needs == | ||
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== References == | == References == | ||
<references /> | <references /> | ||
[[Category: Health]] | |||