Peptide nucleic acids (PNA) demonstrate high affinity to complementary DNA or RNA sequences, and are stable towards enzymatic degradation. These properties make PNA an excellent and robust material for nucleic acid testing, and thus utilized as a class of diagnostic agents. HIV infects millions every year, and one of the limiting factors in curbing the spread of the disease is the capacity to detect the virus in its early stages. In employing PNA as a diagnostic agent, we are able to quantitatively detect the HIV viral load at pertinent levels. HIV-1 gag RNA can be detected using a sandwich hybridization technique, using two complementary probes: a surface PNA probe and a reporter PNA probe. This study focused on increasing the assay sensitivity and optimizing the multivalent architecture. A small library of reporter probes, containing modified monomers, were synthesized to enhance the binding to the RNA target. The multivalent scaffold, containing assorted biotins, was systematically spaced to reduce steric hindrance. These chemical modifications were sequentially added to the reporter PNA probe to evaluate the assay sensitivity, yielding quantitative detection in low femtomolar concentrations.  <br/>

Development of peptide nucleic acids for the optimization of assay sensitivity for quantitative determination of HIV viral load

254th National Meeting (2017)

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