This space represents the ideas, views, opinions, projects and data of researchers within the Aptamer Stream of the Freshman Research Initiative, a program developed at the University of Texas at Austin. These are projects we currently have in the pipeline.
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Nucleic Acid Aptamer Selection against Hcp-6 Surface Protein of Burkholderia pseudomallei for Diagnosis of Melioidosis
Acid Aptamer Selection against Hcp-6 Surface Protein of Burkholderia pseudomallei for Diagnosis of Melioidosis By Dara Goral
Burkholderia pseudomallei is a gram negative bacterium responsible for the disease, melioidosis, which has been the
cause of many fatalities in Southeast Asia and Northern Australia[i]. Melioidosis is a septic disease that often causes abscesses in the spleen and liver[ii].
Because of the resilience of B.
pseudomallei, many antibiotics have not proven effective in reducing
infection and melioidosis is still associated with a high mortality rate of
40%. Most fatalities occur within the first 48 hours of infection and it is
difficult to diagnose this disease in that short amount of time[iii].
approach is introduced that seeks to bind short RNA oligonucleotides to the
Hcp-6 surface protein of Burkholderia pseudomallei. This surface protein is an excellent marker because it is a major virulence determinant for the melioidosis. In the suggested approach, the use of RNA ligands, called
aptamers, prove to be a promising method for binding of Hcp-6 because of successful aptamer selections against many other proteins with various functions. Aptamers are powerful tools in diagnostics and
drug delivery due to their high specificity and affinity for protein targets.
In finding an aptamer with a high affinity for Hcp-6, it could be a crucial
step in the development of a method for diagnosing melioidosis.
Specific Aim 1: Perform SELEX (Systematic Evolution of Ligands by Exponential Enrichment)in order to select an
aptamer to bind to the Hcp-6 surface protein of Burkholderia pseudomallei.
A highly specialized aptamer may be developed that binds with
great affinity for this surface protein allowing for easy identification of
this bacteria in any sample.
Specific Aim 2: Attach a fluorescent tag to the RNA
aptamer and experiment use of detection.
The attachment of a fluorescent tag would allow for easy
detection of Burkholderia pseudomallei
in a given sample. This could expedite the process of diagnosing melioidosis.
Figure 1: The RNA aptamer will bind to
the Hcp-6 surface protein of the bacterium. After this specific aim 1 is
achieved, a fluorescent tag will be attached to the aptamer for detection.
Previous Research: Two rounds of selection have already been completed against this target.
[i]Brett, P. J., D. DeShazer,
and D. E. Woods. (1998) "Burkholderia Thailandensis Sp. Nov., a Burkholderia
Pseudomallei-like Species." International Journal Of Systematic And
Evolutionary Microbiology 48.1: 317-20.
[ii]Currie, Bart J., Dale A.
Fisher, Nicholas M. Anstey, and Susan P. Jacups. (2000) "Melioidosis:
Acute and Chronic Disease, Relapse and Re-activation." Transactions of
the Royal Society of Tropical Medicine and Hygiene 94.3: 301-04.
[iii]Rajchanuvong, A., W.
Chaowagul, Y. Suputtamongkol, M.d. Smith, D.a.b. Dance, and N.j. White. (1995) "A
Prospective Comparison of Co-amoxiclav and the Combination of Chloramphenicol,
Doxycycline, and Co-trimoxazole for the Oral Maintenance Treatment of
Melioidosis." Transactions of the Royal Society of Tropical Medicine
and Hygiene 89.5: 546-49.