Gold aggregating gold: A novel nanoparticle biosensor approach for the direct quantification of hepatitis C virus RNA in clinical samples

The affordable and reliable detection of Hepatitis C Virus (HCV) RNA is a cornerstone in the management and control of infection, affecting approximately 3% of the global population. However, the existing technologies are expensive, labor intensive and time consuming, posing significant limitations to their wide-scale exploitation, particularly in economically deprived populations. Here, we utilized the unique optical and physicochemical properties of gold nanoparticles (AuNPs) to develop a novel assay platform shown to be rapid and robust in sensing and quantifying unamplified HCV RNA in clinical samples. The assay is based on inducing aggregation of citrate AuNPs decorated with a specific nucleic acid probe. Two types of cationic AuNPs, cysteamine and CTAB capped, were compared to achieve maximum assay performance. The technology is simple, rapid, cost effective and quantitative with 93.3% sensitivity, high specificity and detection limit of 4.57 IU/µl. Finally, our data suggest that RNA folding impact the aggregation behavior of the functionalized AuNPs, with broader applications in other nucleic acid detection technologies.

[1]  Xiaohua Huang,et al.  Selective laser photo-thermal therapy of epithelial carcinoma using anti-EGFR antibody conjugated gold nanoparticles. , 2006, Cancer letters.

[2]  T. Pietschmann,et al.  Thermostability of seven hepatitis C virus genotypes in vitro and in vivo , 2013, Journal of viral hepatitis.

[3]  M. Gazouli,et al.  Direct detection of unamplified DNA from pathogenic mycobacteria using DNA-derivatized gold nanoparticles. , 2009, Journal of microbiological methods.

[4]  Chad A Mirkin,et al.  The bio-barcode assay for the detection of protein and nucleic acid targets using DTT-induced ligand exchange , 2006, Nature Protocols.

[5]  Peter Eaton,et al.  Gold nanoparticles for the development of clinical diagnosis methods , 2008, Analytical and bioanalytical chemistry.

[6]  J. Reymond,et al.  Corrigendum: Anti-Microbial Dendrimers against Multidrug-Resistant P. aeruginosa Enhance the Angiogenic Effect of Biological Burn-wound Bandages , 2016, Scientific Reports.

[7]  S. Schrager,et al.  A Brief Clinical Update on Hepatitis C--The Essentials. , 2015, WMJ : official publication of the State Medical Society of Wisconsin.

[8]  Mark A. Atwater,et al.  Extinction coefficient of gold nanoparticles with different sizes and different capping ligands. , 2007, Colloids and surfaces. B, Biointerfaces.

[9]  Paul E. Otto,et al.  Separate Isolation of Genomic DNA and Total RNA from Single Samples Using the SV Total RNA , 1998 .

[10]  Pedro V Baptista,et al.  RNA quantification using gold nanoprobes - application to cancer diagnostics , 2010, Journal of nanobiotechnology.

[11]  Huixiang Li,et al.  Detection of specific sequences in RNA using differential adsorption of single-stranded oligonucleotides on gold nanoparticles. , 2005, Analytical chemistry.

[12]  Reham Atteya,et al.  Topoisomerase-mediated chromosomal break repair: an emerging player in many games , 2015, Nature Reviews Cancer.

[13]  Miguel Larguinho,et al.  Gold nanoprobe-based non-crosslinking hybridization for molecular diagnostics , 2015, Expert review of molecular diagnostics.

[14]  C. Murphy,et al.  Quantitation of metal content in the silver-assisted growth of gold nanorods. , 2006, The journal of physical chemistry. B.

[15]  Kae Sato,et al.  Non-cross-linking gold nanoparticle aggregation as a detection method for single-base substitutions , 2005, Nucleic acids research.

[16]  Aijaz Ahmed,et al.  Update on hepatitis C: Direct-acting antivirals. , 2015, World journal of hepatology.

[17]  M. Farag,et al.  Isoeugenol is a selective potentiator of camptothecin cytotoxicity in vertebrate cells lacking TDP1 , 2016, Scientific Reports.

[18]  D. Gretch,et al.  Molecular diagnostics of hepatitis C virus infection: a systematic review. , 2007, JAMA.

[19]  J. Storhoff,et al.  Selective colorimetric detection of polynucleotides based on the distance-dependent optical properties of gold nanoparticles. , 1997, Science.

[20]  Hongbin Li,et al.  Quantifying thiol–gold interactions towards the efficient strength control , 2014, Nature Communications.

[21]  P. Baptista,et al.  Optimizing Au-nanoprobes for specific sequence discrimination. , 2010, Colloids and surfaces. B, Biointerfaces.

[22]  H. Azzazy,et al.  Detection of unamplified HCV RNA in serum using a novel two metallic nanoparticle platform , 2014, Clinical chemistry and laboratory medicine.

[23]  J. Turkevich,et al.  Colloidal gold. Part II , 1985 .

[24]  Jian Wang,et al.  A gold nanoparticles-based colorimetric assay for alkaline phosphatase detection with tunable dynamic range. , 2013, Biosensors & bioelectronics.

[25]  J. Turkevich,et al.  Colloidal gold. Part I , 1985 .

[26]  Huixiang Li,et al.  Colorimetric detection of DNA sequences based on electrostatic interactions with unmodified gold nanoparticles. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[27]  Mizuo Maeda,et al.  Rapid aggregation of gold nanoparticles induced by non-cross-linking DNA hybridization. , 2003, Journal of the American Chemical Society.

[28]  C. Liao,et al.  Epigenetic changes in histone acetylation underpin resistance to the topoisomerase I inhibitor irinotecan , 2016, Nucleic acids research.

[29]  L. Liz‐Marzán,et al.  A "Tips and Tricks" Practical Guide to the Synthesis of Gold Nanorods. , 2015, The journal of physical chemistry letters.

[30]  Huixiang Li,et al.  Label-free colorimetric detection of specific sequences in genomic DNA amplified by the polymerase chain reaction. , 2004, Journal of the American Chemical Society.

[31]  Sang J. Chung,et al.  An operationally simple colorimetric assay of hyaluronidase activity using cationic gold nanoparticles. , 2009, The Analyst.

[32]  L. Abu-Raddad,et al.  Evidence of intense ongoing endemic transmission of hepatitis C virus in Egypt , 2010, Proceedings of the National Academy of Sciences.

[33]  G. Dore,et al.  Epidemiology and natural history of HCV infection , 2013, Nature Reviews Gastroenterology &Hepatology.

[34]  H. Azzazy,et al.  Direct detection of unamplified hepatoma upregulated protein RNA in urine using gold nanoparticles for bladder cancer diagnosis. , 2014, Clinical biochemistry.

[35]  P. Jain,et al.  Gold nanoparticles: interesting optical properties and recent applications in cancer diagnostics and therapy. , 2007, Nanomedicine.

[36]  G. Dawson,et al.  Role of Serologic and Molecular Diagnostic Assays in Identification and Management of Hepatitis C Virus Infection , 2015, Journal of Clinical Microbiology.

[37]  Chad A Mirkin,et al.  Maximizing DNA loading on a range of gold nanoparticle sizes. , 2006, Analytical chemistry.

[38]  R. Kamel AASLD 2016 Recommendations for Testing, Managing, and Treating Hepatitis C , 2017 .

[39]  S. Goueli,et al.  Positional effect of mutations in 5'UTR of hepatitis C virus 4a on patients' response to therapy. , 2009, World journal of gastroenterology.

[40]  Keng-Shiang Huang,et al.  An electroporation microchip system for the transfection of zebrafish embryos using quantum dots and GFP genes for evaluation , 2007, Biomedical microdevices.