Non-instrumented Nucleic Acid Extraction Technology for Enabling Point-of-Care Assays to Assess HIV Resistance

In 2010, approximately 35 million adults and children were living with HIV and 2.7 million people were newly infected (WHO). If a patient has the mutant HIV pol gene, alternative treatment and constant monitoring are required. An oligonucleotide ligation assay (OLA) was developed that can use a whole blood sample or nucleic acids (NA) to test for the mutant HIV pol gene. However, a key challenge facing the current OLA for downstream PCR is the extraction of NAs because the extraction requires a lengthy protocol, using instrumentation unavailable in low resource settings, where 75% of those who have HIV live. To address this issue, we are developing ways to capture NAs without instrumentation through the use of stimuli-responsive polymers. Stimuli-responsive polymers are reagents capable of responding to specific changes within their environments such as temperature or pH variations. Magnetic nanoparticles (mNPs) can be utilized in conjunction with these polymers for biomolecule separation by applying a stimulus and a magnetic field. Our NA extraction strategy utilizes stimuli-responsive cationic polymers and mNPs to complex and magnetically separate NAs successfully. Electrophoresis gels were used to characterize the successful complexation efficiency of the polymer and NA at ideal conditions. It was observed that the complexation efficiency was dose responsive. As this research continues, it will address health disparities through its aim of being accessible globally in low resource settings, without using instrumentation, and to be used in point-of-care diagnostic tests for the HIV pol gene to provide correct treatment methods for patients.