Nucleic Acid Aptamer Selection Against Hcp-6 for Diagnosis and Detection of Burkholderia pseudomallei

Nucleic Acid Aptamer Selection Against Hcp-6 for Diagnosis and Detection of Burkholderia pseudomallei           

Armin Nourani

         Throughout many areas of Southeast Asia and surrounding regions, Burkholderia pseudomallei and several related strains have been responsible for an uprising in a disease known as melioidosis [1]. This disease has a high mortality rate, reaching nearly 40-80% in areas such as Thailand where treatment is low and the bacteria has the ability to spread rapidly through many water sources [2]. Treatment for this disease is possible and more effective when begun earlier; however, early detection is lacking due to the current diagnosis method of culturing, which is both timely and costly [2]. Thus, poor diagnosis is a leading factor for such high mortality rates [3][4].

The current diagnosis method could be vastly improved through the use of an aptamer selection against a protein known as Hcp-6. It is a surface protein involved in the secretion system in B. pseudomallei, the strain most responsible for melioidosis, and is an ideal target for an aptamer selection due to its prominent location on the exterior of the bacteria. Furthermore, an aptamer for Hcp-6 could also be used for detection of contamination in water sources, which would be exponentially useful in limiting the spreading of the bacteria to others.

Figure 1: This image represents what is hoped to be gained from doing an aptamer selection against Hcp-6. By combining both a highly selective aptamer with a visual signal, recognition of B. pseudomallei can be achieved and applied in diagnostics and other contamination tests.

Specific Aim 1: Develop a selective RNA aptamer against Hcp-6.  In order to find an aptamer used for the recognition of this bacteria, it is essential that it is highly specific and can bind well to the volatile protein for optimum use.
Specific Aim 2: Develop a real-world method of using an Hcp-6 aptamer for its various purposes. Once an aptamer has been found, it is necessary to develop a detection method for its binding to Hcp-6 and an a means of adapting that to an easily reproducible, cost effective tool to be used for both human samples and water samples.

Ordering Information: Hcp-6 is provided through the Katy Brown Lab, and is thus free of charge. It is primarily obtained through Dr. Omar Qazi.

References:

[1] Wuthiekanun, V, Peacock, S.J. (2006). “Management of Melioidosis.” Expert Rev Anti Infect Ther. 4(3):445-55.

[2] Warner, J.M., Pelowa, D.B., Currie, B.J., Hirst, R.G. (2007). “Melioidosis in a rural community of Western Province, Papua New Guinea.” Trans R Soc Trop Med Hyg. 101(8):809-13.

[3] Walsh, A.L., Wuthiekanun, V. (1996). “The Laboratory Diagnosis of  Melioidosis.” Br J Biomed Sci. 53(4):249-53.

[4] Limmathurotsakul, D., Wuthiekanun, V., Chierakul, W., Cheng, A.C., Maharjan, B., Chaowagul, W., White, N.J., Day, N.P.J., Peacock, S.J. (2005). “Role and Significance of Quantitative Urine Cultures in Diagnosis of Melioidosis.” J Clin Microbiol. 43(5): 2274–2276.

Proposal: Hcp-6 Proposal
First Progress Report: Hcp-6 First Progress Report
Second Progress Report: Hcp-6 Second Progress Report
Final Progress Report: Hcp-6 Final Progress Report