Hepatitis C virus (HCV) infection is the primary etiologic agent of liver cirrhosis or hepatocellular carcinoma. In the United States (U.S.) alone, &gt;4M persons are found to be HCV antibody positive and &gt;2M persons HCV RNA positive, consistent with active infection. Current POC tests only screens for anti-HCV antibodies or HCV antigens, and requires the patient to undergo separate testing for confirmatory HCV RNA and genotyping to determine active infection. Herein, we present a novel nanotechnology-enabled biochemical assay for multiplexed detection of active HCV RNA integrated with a rapid, point-of-care (POC) genotyping platform. To achieve molecular targeting, antisense oligonucleotides (ASOs) have been designed <i>in silico</i> to target different segments of HCV genome. Two ASOs have been designed targeting a conserved HCV RNA sequence, NS-5b, present among all the genotypes. For genotyping, we have targeted the 5’-UTR and 3’-UTR of different HCV genotypes. The ASO-AuNPs agglomerated in presence of its target RNA sequence and demonstrated a change in their surface plasmon resonance (SPR). The agglomeration of AuNPs resulted in a change in color, enabling visual naked-eye detection of HCV. Although a few POC biosensors have been proposed very recently for HCV, this will be the first study to develop an integrated multiplexed assay for early detection and genotyping both within a same platform, fulfilling an unmet global need. The technique is capable of detecting the viral load directly from plasma and works without a requirement of RNA isolation, making it a true POC. We anticipate that our sensor can detect the specific target nucleic acid sequences without signal cross talk with a detection limit of ~26 IU/liter and a time of response (TOR) of around 5 mins. We expect that the current point-of-care assay from whole plasma would allow patients to receive their result in a single visit and allow for rapid care planning and management.

Selective naked-eye point-of-care detection of HCV genotypes mediated by new antisense oligonucleotide capped plasmonic nanoparticles

ACS Spring Meeting 2021

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