Virus detection and quantification using electrical parameters

Here we identify and quantitate two similar viruses, human and feline immunodeficiency viruses (HIV and FIV), suspended in a liquid medium without labeling, using a semiconductor technique. The virus count was estimated by calculating the impurities inside a defined volume by observing the change in electrical parameters. Empirically, the virus count was similar to the absolute value of the ratio of the change of the virus suspension dopant concentration relative to the mock dopant over the change in virus suspension Debye volume relative to mock Debye volume. The virus type was identified by constructing a concentration-mobility relationship which is unique for each kind of virus, allowing for a fast (within minutes) and label-free virus quantification and identification. For validation, the HIV and FIV virus preparations were further quantified by a biochemical technique and the results obtained by both approaches corroborated well. We further demonstrate that the electrical technique could be applied to accurately measure and characterize silica nanoparticles that resemble the virus particles in size. Based on these results, we anticipate our present approach to be a starting point towards establishing the foundation for label-free electrical-based identification and quantification of an unlimited number of viruses and other nano-sized particles.

[1]  A. Kelso,et al.  A comparison of rapid point-of-care tests for the detection of avian influenza A(H7N9) virus, 2013. , 2013, Euro surveillance : bulletin Europeen sur les maladies transmissibles = European communicable disease bulletin.

[2]  A. Heck,et al.  Interrogating viral capsid assembly with ion mobility-mass spectrometry. , 2011, Nature chemistry.

[3]  J. McKimm-Breschkin,et al.  Plaque formation assay for human parainfluenza virus type 1. , 2011, Biological & pharmaceutical bulletin.

[4]  S. Gan,et al.  Enzyme immunoassay and enzyme-linked immunosorbent assay. , 2013, The Journal of investigative dermatology.

[5]  J. Slater,et al.  Competition enzyme‐linked immunosorbant assay (ELISA) can be a sensitive method for the specific detection of small quantities of allergen in a complex mixture , 2006, Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology.

[6]  P. Kellam,et al.  Full Genome Virus Detection in Fecal Samples Using Sensitive Nucleic Acid Preparation, Deep Sequencing, and a Novel Iterative Sequence Classification Algorithm , 2014, PloS one.

[7]  P. Roingeard Viral detection by electron microscopy: past, present and future , 2008, Biology of the cell.

[8]  G. Yarrington Molecular Cell Biology , 1987, The Yale Journal of Biology and Medicine.

[9]  L. Rychlewski,et al.  Alphaherpesvirinae and Gammaherpesvirinae glycoprotein L and CMV UL130 originate from chemokines , 2013, Virology Journal.

[10]  A. Nitsche,et al.  Real-time PCR in virology. , 2002, Nucleic acids research.

[11]  D. Holdstock Past, present--and future? , 2005, Medicine, conflict, and survival.

[12]  P E Klapper,et al.  Multiplex PCR: Optimization and Application in Diagnostic Virology , 2000, Clinical Microbiology Reviews.

[13]  S. Sze,et al.  Physics of Semiconductor Devices: Sze/Physics , 2006 .

[14]  G. Pauli,et al.  Determination of the size of HIV using adenovirus type 2 as an internal length marker. , 1994, Journal of virological methods.

[15]  T. Graule,et al.  Isoelectric points of viruses , 2009, Journal of applied microbiology.

[16]  Oumar Faye,et al.  Quantitative real-time PCR detection of Zika virus and evaluation with field-caught Mosquitoes , 2013, Virology Journal.

[17]  P. Klapper,et al.  Multiplex PCR: Optimization and Application in Diagnostic Virology , 2000, Clinical Microbiology Reviews.

[18]  Yao Xiao,et al.  Two types of nanoparticle-based bio-barcode amplification assays to detect HIV-1 p24 antigen , 2012, Virology Journal.

[19]  Lee K. Rhea,et al.  How Much Is Enough? Minimal Responses of Water Quality and Stream Biota to Partial Retrofit Stormwater Management in a Suburban Neighborhood , 2014, PloS one.

[20]  E. Luzzi,et al.  A Competitive Enzyme-Linked Immunosorbent Assay for Measuring the Levels of Serum Antibody to Haemophilus influenzae Type b , 1998, Clinical Diagnostic Laboratory Immunology.

[21]  F. Mustafa,et al.  Optimal Packaging of FIV Genomic RNA Depends upon a Conserved Long-range Interaction and a Palindromic Sequence within gag , 2010, Journal of molecular biology.

[22]  Julio C. B. Ferreira,et al.  Protein Quality Control Disruption by PKCβII in Heart Failure; Rescue by the Selective PKCβII Inhibitor, βIIV5-3 , 2012, PloS one.

[23]  B. Verhasselt,et al.  Quantification of Reverse Transcriptase Activity by Real-Time PCR as a Fast and Accurate Method for Titration of HIV, Lenti- and Retroviral Vectors , 2012, PloS one.

[24]  G. Sauer,et al.  Identification of Virus-Induced Proteins in Cells Productively Infected with Simian Virus 40 , 1972, Journal of virology.

[25]  J. Damen,et al.  Hematopoietic colony-forming cell assays. , 2007, Methods in molecular biology.

[26]  J. Phair,et al.  Detection of HIV-1 p24 Gag in plasma by a nanoparticle-based bio-barcode-amplification method. , 2008, Nanomedicine.

[27]  T. Rizvi,et al.  Primate and Feline Lentivirus Vector RNA Packaging and Propagation by Heterologous Lentivirus Virions , 2001, Journal of Virology.

[28]  H. Grubin The physics of semiconductor devices , 1979, IEEE Journal of Quantum Electronics.

[29]  Gengfeng Zheng,et al.  Electrical detection of single viruses. , 2004, Proceedings of the National Academy of Sciences of the United States of America.