MicroRNA Purification for Diagnostic Development (2024)

Undergraduate: Anay Shah

Faculty Advisor: Dr. Adam Akerman
Department: Cardiothoracic Surgery

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Aortic aneurysms are abnormal bulges in the walls of the aorta and pose a threat to humans because of their possibility of rupture. Increasing vessel size leads to an increased pressure that could indicate a higher likelihood for an aneurysm rupture. This could lead to false lumen, internal bleeding, and other cardiothoracic complications. Current testing for aortic aneurysms is expensive and oftentimes aortic aneurysms are exposed accidentally through imaging techniques for other conditions. Because of this, developing a cheap and easy way to test for aortic aneurysms is important. Because certain microRNAs are secreted into the bloodstream with aortic aneurysm growth, like mir-1339 and mir-133b, quantifying those microRNAs can be a method for diagnostic testing associated with a blood test. The first step of microRNA quantification comes from purifying microRNA sequences from plasma samples. Charge purification methods were used for isolating the 22 nucleotide sequences from the plasma samples. Charge purification calls for washing the plasma samples with buffers, the addition of ethanol to reverse the charge of the nucleic acid, and eluting the microRNA sequences in RNAse free water. This eluted sample is inputted for Qubit Analysis in which the amount of microRNAs can be calculated in ng/ul. Future work involves standardizing the concentrations of all the samples, selecting exogenous and endogenous controls, cDNA generation, and digital droplet PCR. The end goal is to perform an ROC curve to test if microRNAs can be used as a diagnostic test for aortic aneurysms and implement the blood test.