Design and characterisation of high-affinity aptamers for detecting HIV integrase

Area of Science:

• Biotechnology
• Molecular Diagnostics
• Virology

Background:

• 39.9 million people globally are infected with HIV, with 1.3 million new infections annually.
• Current diagnostic tests cannot detect HIV within the first 10 days of infection, delaying crucial treatment.
• Aptamers offer a promising alternative to antibodies for specific protein recognition in diagnostic assays.

Purpose of the Study:

• To develop novel aptamers for early detection of diverse human immunodeficiency virus (HIV) variants.
• To identify aptamer combinations for sensitive and specific detection of HIV integrase (IN).

Main Methods:

• Bioinformatic analysis of over 20,000 HIV-1 and HIV-2 integrase sequences to identify conserved targets.
• Selection and characterization of aptamers targeting a highly conserved peptide in HIV integrase.
• Enzyme-Linked Oligonucleotide Assays (ELONA) to evaluate aptamer affinity, sensitivity, and specificity.

Main Results:

• Three aptamers were selected targeting a conserved peptide in HIV integrase across numerous variants.
• The most effective aptamer combination detected HIV at levels as low as 3.65 log HIV cp/mL.
• The aptamers demonstrated high specificity, not reacting with negative controls or other viruses.

Conclusions:

• Developed anti-HIV integrase aptamers show potential for ultrasensitive nanotechnologies.
• These aptamers could enable early HIV diagnosis from the first day of infection across all variants.
• Future aptamer-based biosensors may revolutionize HIV diagnostics, irrespective of patient demographics or infection stage.