Single virus detection by means of atomic force microscopy in combination with advanced image analysis.

In the present contribution virions of five different virus species, namely Varicella-zoster virus, Porcine teschovirus, Tobacco mosaic virus, Coliphage M13 and Enterobacteria phage PsP3, are investigated using atomic force microscopy (AFM). From the resulting height images quantitative features like maximal height, area and volume of the viruses could be extracted and compared to reference values. Subsequently, these features were accompanied by image moments, which quantify the morphology of the virions. Both types of features could be utilized for an automatic discrimination of the five virus species. The accuracy of this classification model was 96.8%. Thus, a virus detection on a single-particle level using AFM images is possible. Due to the application of advanced image analysis the morphology could be quantified and used for further analysis. Here, an automatic recognition by means of a classification model could be achieved in a reliable and objective manner.

[1]  A. McPherson,et al.  Imaging of viruses by atomic force microscopy. , 2001, The Journal of general virology.

[2]  Chin-Cheng Huang,et al.  The role of porcine teschovirus in causing diseases in endemically infected pigs. , 2012, Veterinary microbiology.

[3]  A. Kuhn,et al.  Kinetics of filamentous phage assembly. , 2010, Physical biology.

[4]  P. Nazarov,et al.  Viruses: incredible nanomachines. New advances with filamentous phages , 2009, European Biophysics Journal.

[5]  D. Anselmetti,et al.  Apertureless scanning near-field optical microscopy of sparsely labeled tobacco mosaic viruses and the intermediate filament desmin , 2013, Beilstein journal of nanotechnology.

[6]  Bengt Jönsson,et al.  Internal DNA pressure modifies stability of WT phage , 2007, Proceedings of the National Academy of Sciences.

[7]  M. Ogilvie Molecular techniques should not now replace cell culture in diagnostic virology laboratories , 2001, Reviews in medical virology.

[8]  Scanning Tunneling and Atomic Force Microscopy of T4 Bacteriophage and Tobacco Mosaic Virus , 1993 .

[9]  A. Arvin,et al.  Varicella in the fetus and newborn. , 2009, Seminars in fetal & neonatal medicine.

[10]  Marc Eloit,et al.  The human virome: new tools and concepts , 2013, Trends in Microbiology.

[11]  L. Makowski Phage display: structure, assembly and engineering of filamentous bacteriophage M13 , 1994 .

[12]  E. Lofgren,et al.  Influenza Seasonality: Underlying Causes and Modeling Theories , 2006, Journal of Virology.

[13]  C. Grose,et al.  Computer modeling of prototypic and aberrant nucleocapsids of varicella-zoster virus. , 1995, Virology.

[14]  S. Nii Electron microscopic study on the development of herpesviruses. , 1992, Journal of electron microscopy.

[15]  Chuanbin Mao,et al.  Die Anwendung von Viren in Chemo‐ und Biosensoren , 2009 .

[16]  H. Gelderblom,et al.  Electron microscopy for rapid diagnosis of infectious agents in emergent situations. , 2003, Emerging infectious diseases.

[17]  J. Schramlová,et al.  The role of electron microscopy in the rapid diagnosis of viral infections — review , 2010, Folia Microbiologica.

[18]  P. Haris,et al.  FT-IR spectroscopy of the major coat protein of M13 and Pf1 in the phage and reconstituted into phospholipid systems. , 1995, Biochemistry.

[19]  A. McPherson,et al.  Atomic Force Microscopy in Imaging of Viruses and Virus-Infected Cells , 2011, Microbiology and Molecular Reviews.

[20]  J. Hsu,et al.  Surface rigidity change of Escherichia coli after filamentous bacteriophage infection. , 2009, Langmuir : the ACS journal of surfaces and colloids.

[21]  C. Grose,et al.  Egress of varicella-zoster virus from the melanoma cell: a tropism for the melanocyte , 1995, Journal of virology.

[22]  Jürgen Popp,et al.  Raman to the limit: tip‐enhanced Raman spectroscopic investigations of a single tobacco mosaic virus , 2009 .

[23]  A. Bressan,et al.  Immunofluorescence localisation of Banana bunchy top virus (family Nanoviridae) within the aphid vector, Pentalonia nigronervosa, suggests a virus tropism distinct from aphid-transmitted luteoviruses. , 2011, Virus research.

[24]  S. Larson,et al.  Structural transitions of satellite tobacco mosaic virus particles. , 2001, Virology.

[25]  Miguel A R B Castanho,et al.  An overview of the biophysical applications of atomic force microscopy. , 2003, Biophysical chemistry.

[26]  S. Okitsu,et al.  Anti‐norovirus polyclonal antibody and its potential for development of an antigen‐ELISA , 2007, Journal of medical virology.

[27]  H. Shu,et al.  Image description with generalized pseudo-Zernike moments. , 2007, Journal of the Optical Society of America. A, Optics, image science, and vision.

[28]  G. Zellnig,et al.  Rapid diagnosis of plant virus diseases by transmission electron microscopy. , 2009, Journal of virological methods.

[29]  G. Borgefors,et al.  Segmentation of virus particle candidates in transmission electron microscopy images , 2012, Journal of microscopy.

[30]  I. Yaminsky,et al.  Atomic force microscopy investigation of phage infection of bacteria. , 2008, Langmuir : the ACS journal of surfaces and colloids.

[31]  J. Sørensen,et al.  Large Pseudomonas phages isolated from barley rhizosphere , 1995 .

[32]  An atomic force microscopy investigation of cyanophage structure. , 2012, Micron.

[33]  P. Descouts,et al.  Preparation of isolated biomolecules for SFM observations: T4 bacteriophage as a test sample. , 1993, Biophysical journal.

[34]  William N. Venables,et al.  Modern Applied Statistics with S , 2010 .

[35]  K. Scholthof Tobacco mosaic virus: a model system for plant biology. , 2004, Annual review of phytopathology.

[36]  Jürgen Popp,et al.  Automated seeding-based nuclei segmentation in nonlinear optical microscopy. , 2013, Applied optics.

[37]  M. Williams,et al.  Morphology of varicella (chicken pox) virus. , 1962, Virology.

[38]  S. Chiu,et al.  Multiple models of porcine teschovirus pathogenesis in endemically infected pigs. , 2014, Veterinary microbiology.

[39]  Jean-Michel Claverie,et al.  Pandoraviruses: Amoeba Viruses with Genomes Up to 2.5 Mb Reaching That of Parasitic Eukaryotes , 2013, Science.

[40]  K. Nakamura,et al.  Experimental Teschovirus Encephalomyelitis in Gnotobiotic Pigs , 2013, Journal of Comparative Pathology.

[41]  L. Kaiser,et al.  Point-Counterpoint: Is the Era of Viral Culture Over in the Clinical Microbiology Laboratory? , 2012, Journal of Clinical Microbiology.

[42]  Jean-Luc Pellequer,et al.  Nanoscale structural features determined by AFM for single virus particles. , 2013, Nanoscale.

[43]  Andrew J. Senesi,et al.  Self-assembly of rodlike virus to superlattices. , 2013, Langmuir : the ACS journal of surfaces and colloids.

[44]  D. Leland,et al.  Role of Cell Culture for Virus Detection in the Age of Technology , 2007, Clinical Microbiology Reviews.

[45]  Norbert Bannert,et al.  Evaluation of tip-enhanced Raman spectroscopy for characterizing different virus strains. , 2011, The Analyst.

[46]  Chee-Way Chong,et al.  The scale invariants of pseudo-Zernike moments , 2003, Pattern Analysis & Applications.

[47]  J. Flusser,et al.  Moments and Moment Invariants in Pattern Recognition , 2009 .

[48]  Mitchel J Doktycz,et al.  Atomic force microscopy of biological samples. , 2010, Wiley interdisciplinary reviews. Nanomedicine and nanobiotechnology.

[49]  I. Nicholls,et al.  Filamentous bacteriophage stability in non-aqueous media. , 2001, Chemistry & biology.