Dustin Taylor - Link to Proposal

Dustin Taylor

Here's my updated abstract.

Aptamer Selection Against Burkholderia pseudomallei Fimbria Protein for Efficient Diagnosis of Melioidosis

Dustin Taylor – Fall 2011

September 16^th, 2011

N58 RNA – B. pseudomallei fimbriae

Burkholderia pseudomallei is a gram negative bacterium that causes the disease melioidosis, marked by the presence of joint pain, cough, skin infections, lung nodules, and pneumonia. It has a niche in soil and surface water, and as a consequence is endemic in Southeast Asian populations where contact with these environments is essentially inevitable.⁴ It is most commonly acquired through a break in the skin, or by inhaling the aerosolized form of the bacterium.

Most current diagnostics include a complete screening involving numerous cultures and a throat swab; this is very time consuming, and only adds to the 50% mortality rate. Additionally, rapid discovery methods, such as antibody and antigen detection, have relatively low sensitivities averaging around 70%, and as a consequence are not highly trusted.³ The goal of this experiment, then, was to develop an aptamer to more efficiently reveal the presence of a B. pseudomallei infection.

Fimbriae from this particular bacterium have been isolated in order to more easily develop aptamer binders. The specific aim was to select for RNA fragments that demonstrate a high binding affinity for the fimbriae; as this could lead to the eventual development of aptamers for faster diagnostics of the presence of this bacterium in a particular sample.

Figure 1: Illustration of aptamer binding. The specific aim for this selection is to find an aptamer that will bind a bacterial fimbria and provide a quicker diagnostic test for the presence of B. pseudomallei.

The fimbriae proteins have been procured by Kate McCaul of Dr. Kate Brown’s lab. The only remaining step with cost is to functionalize the proteins with biotin. This cost is unknown as the amount of biotin used is target-specific, and has not been recorded.

Here's a link to my proposal.

Here is a link to my first progress report.

Here is a link to my final manuscript.