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Detection of unamplified HCV RNA in serum using a novel two metallic nanoparticle platform

  • Sherif M. Shawky , Bassem S. Guirgis and Hassan M.E. Azzazy EMAIL logo
Published/Copyright: October 25, 2013
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Clinical Chemistry and Laboratory Medicine
From the journal Clinical Chemistry and Laboratory Medicine Volume 52 Issue 4

Abstract

Background: The unique properties of metallic nanoparticles have enabled their utilization in biosensing applications. A novel assay for the detection of hepatitis C virus (HCV) RNA in serum specimens has been developed using magnetic nanoparticles and unmodified cationic gold nanoparticles (AuNPs).

Methods: HCV RNA was extracted using magnetic nanoparticles functionalized with an oligonucleotide specific to HCV RNA. Extracted RNA is reacted with oligonucleotide sequence specific for HCV RNA in presence of unmodified cationic AuNPs. In positive samples, AuNPs are aligned onto the phosphate backbone of the RNA and their aggregation changes the solution color from red to blue. In the absence of target, solution color remains red. The assay has been tested on 50 serum clinical samples (25 HCV positive and 25 controls).

Results: The dual nanoparticles assay detected HCV RNA in serum and generated comparable results to real-time PCR. The assay had specificity and a sensitivity of 96% and 96.5%, respectively, and a detection limit of 15 IU/mL.

Conclusions: The developed colorimetric dual nanoparticles HCV RNA assay is simple and inexpensive and can be used for rapid detection of unamplified HCV RNA in serum. Similar sensing platforms can be developed to detect other nucleic acid targets.

Keywords: cationic AuNPs; hepatitis C virus (HCV); magnetic nanoparticles; nucleic acid tests (NAT); qualitative HCV RNA assay
Corresponding author: Hassan M.E. Azzazy, PhD, Department of Chemistry, School of Sciences & Engineering, The American University in Cairo, P.O. Box 74 New Cairo, 11835 Egypt, Phone: +20 1000565727, E-mail:
aSherif M. Shawky and Bassem S. Guirgis contributed equally to this work.

Acknowledgments

The authors acknowledge Mr. Kamel Eid (YJ-STRC) for preparing the magnetic nanoparticles.

Conflict of interest statement

Authors’ conflict of interest disclosure: The authors stated that there are no conflicts of interest regarding the publication of this article. Research funding played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

Research funding: This work has been funded by a grant from YJ-STRC (AUC) to Dr. Hassan Azzazy.

Employment or leadership: Authors of this paper have submitted a patent on the use of nanoparticles for detection of HCV RNA in clinical specimens. Dr. Azzazy is the President and Chief Technology Officer of D-Kimia, a LLC, which develops HCV nanodiagnostic assays.

Honorarium: None declared.

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Received: 2013-7-9
Accepted: 2013-9-27
Published Online: 2013-10-25
Published in Print: 2014-04-01

©2014 by Walter de Gruyter Berlin/Boston

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https://doi.org/10.1515/cclm-2013-0521
Keywords for this article
cationic AuNPs; hepatitis C virus (HCV); magnetic nanoparticles; nucleic acid tests (NAT); qualitative HCV RNA assay

Articles in the same Issue

  1. Masthead
  2. Masthead
  3. Editorials
  4. Inflammatory bowel diseases: where we are and where we should go
  5. Thrombophilia testing. Useful or hype?
  6. Reviews
  7. Inflammatory bowel diseases: from pathogenesis to laboratory testing
  8. Crohn’s disease specific pancreatic antibodies: clinical and pathophysiological challenges
  9. Point/Counterpoint
  10. The utility of thrombophilia testing
  11. The futility of thrombophilia testing
  12. Genetics and Molecular Diagnostics
  13. Identification of an 18 bp deletion in the TWIST1 gene by CO-amplification at lower denaturation temperature-PCR (COLD-PCR) for non-invasive prenatal diagnosis of craniosynostosis: first case report
  14. General Clinical Chemistry and Laboratory Medicine
  15. Agreement of seven 25-hydroxy vitamin D3 immunoassays and three high performance liquid chromatography methods with liquid chromatography tandem mass spectrometry
  16. First trimester pregnancy-associated plasma protein A and human chorionic gonadotropin-beta in early and late pre-eclampsia
  17. S100B blood level measurement to exclude cerebral lesions after minor head injury: the multicenter STIC-S100 French study
  18. NGAL, L-FABP, and KIM-1 in comparison to established markers of renal dysfunction
  19. Value-added reporting of antinuclear antibody testing by automated indirect immunofluorescence analysis
  20. Cardiovascular Diseases
  21. A fit-for-purpose approach to analytical sensitivity applied to a cardiac troponin assay: time to escape the ‘highly-sensitive’ trap
  22. Diabetes
  23. A simple and precise method for direct measurement of fractional esterification rate of high density lipoprotein cholesterol by high performance liquid chromatography
  24. Infectious Diseases
  25. Detection of unamplified HCV RNA in serum using a novel two metallic nanoparticle platform
  26. Increased plasma arginase activity in human sepsis: association with increased circulating neutrophils
  27. Corrigendum
  28. The relationship between estimated average glucose and fasting plasma glucose
  29. Letter to the Editors
  30. The obsessive-compulsory clinical pathologist
  31. Prism effect of specimen receiving – generation of fundamentals for the smooth progress of analytical processing
  32. Preanalytical errors: the professionals’ perspective
  33. Corrected reports in laboratory medicine in a Chinese university hospital for 3 years
  34. Comparison of an enzymatic assay with liquid chromatography-pulsed amperometric detection for the determination of lactulose and mannitol in urine of healthy subjects and patients with active celiac disease
  35. Effect of freezing-thawing process on neuron specific enolase concentration in severe traumatic brain injury sera samples
  36. Undetected creatinine levels
  37. The effect of centrifugation on three urine protein assays: benzethonium chloride, benzalkonium chloride and pyrogallol red
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