Nucleic Acid Aptamer Selection Against Burkholderia Pseudomallei for the diagnostic of Meliodosis

Nucleic Acid Aptamer Selection Against Burkholderia Pseudomallei for the diagnostic of Meliodosis
by Fidel Fernandez

Burkholderia Pseudomallei is the bacterium responsible for causing the disease Meliodosis. Burkholderia Pseudomallei infects primarily Southeast Asia and Northern Australia and is common among rice field workers. With four different types of infections caused by Burkholderia Pseudomallei, and a Category B bioterrorism agent listing by the Center of Disease Control (CDC), the need for a better diagnostic test is needed. The current diagnostic test is the simple yet effective blood culture. The problem however, with the blood culture, is that this test may take anywhere from 24-48 hours to cultivate and the time spent diagnosing the infection could be better spent on treating the infection.

The four types of the infections include the localized, pulmonary, septicemia, and chronic forms. The localized form causes swollen lymph nodes along with the development of ulcers. Pulmonary abscesses and Pneumonia are both common symptoms associated with the pulmonary form of Meliodosis.  In the septicemia form, 90% of patients die within the first 24-48 hours due to splenic, cutaneous, and hepatic difficulties. Multiple abscesses form on the liver, spleen, skin, and muscle tissues, which become more reactive with passing years.

An aptamer against the flagella of Burkholderia Pseudomallei would be beneficial because of the time it takes to diagnose the disease. As listed above, the septicemia form of the disease can kill the infected within the first 24-48 hours, which is the same amount of time that it takes to diagnose the disease with the current diagnostic tool, the blood culture. If an aptamer against the flagella of Burkholderia Pseudomallei was found, then the time needed to diagnose the infection could be reduced to only a few minutes.

                                                                            In Figure 1, located to the left, a flagella is looked at more closely

and located in the filament of the flagella, proteins are found that are the binding site for the potential aptamer. An aptamer would bind to those proteins on the filament of the flagella.

Specific Aim:

To isolate an aptamer against the Burkholderia Pseudomallei Flagella for the purpose to diagnostics.

Aptamer Selection Target

Target Name (full name): BPSL3319 Flagellin (native)

Abbreviated Name(s): FLA

Date Obtained: June 2013

Acquired Target from: Dr. Annie Gnanam, Dr. Katy Brown’s lab

Functionalized/tagged: No

Molecular Weight: 39,300 g/mol 

Full Proposal:
https://docs.google.com/file/d/0B87nMSoAVDzpRzhzQWNuY2dxZU0/edit?usp=sharing

First Progress Report:
https://docs.google.com/file/d/0B87nMSoAVDzpekVjaFlHRmt0Yk0/edit?usp=sharing

Second Progress Report:
https://drive.google.com/file/d/0B87nMSoAVDzpX1Z1OWl2UmZPb2s/edit?usp=sharing

Final Report:
https://drive.google.com/file/d/0B87nMSoAVDzpQjZqeVBzNlJDWWs/edit?usp=sharing

References:

"Burkholderia Pseudomallei." Burkholderia Pseudomallei. Pathema, 18 May 2006. Web. 02 Sept. 2013. 

"Meliodosis." Centers for Disease Control and Prevention. Centers for Disease Control and Prevention, 26 Jan. 2012. Web. 02 Sept. 2013. 

MLST. "Burkholderia Pseudomallei - Organismal Information." Burkholderia Pseudomallei - Organismal Information. MLST, n.d. Web. 02 Sept. 2013.