Digital sensing and sizing of vesicular stomatitis virus pseudotypes in complex media: a model for Ebola and Marburg detection.
Rapid, sensitive, and direct label-free capture and characterization of nanoparticles from complex media such as blood or serum will broadly impact medicine and the life sciences. We demonstrate identification of virus particles in complex samples for replication-competent wild-type vesicular stomatitis virus (VSV), defective VSV, and Ebola- and Marburg-pseudotyped VSV with high sensitivity and specificity. Size discrimination of the imaged nanoparticles (virions) allows differentiation between modified viruses having different genome lengths and facilitates a reduction in the counting of nonspecifically bound particles to achieve a limit-of-detection (LOD) of 5 × 10(3) pfu/mL for the Ebola and Marburg VSV pseudotypes. We demonstrate the simultaneous detection of multiple viruses in a single sample (composed of serum or whole blood) for screening applications and uncompromised detection capabilities in samples contaminated with high levels of bacteria. By employing affinity-based capture, size discrimination, and a "digital" detection scheme to count single virus particles, we show that a robust and sensitive virus/nanoparticle sensing assay can be established for targets in complex samples. The nanoparticle microscopy system is termed the Single Particle Interferometric Reflectance Imaging Sensor (SP-IRIS) and is capable of high-throughput and rapid sizing of large numbers of biological nanoparticles on an antibody microarray for research and diagnostic applications.
Electrical & Computer Engineering Dept., Boston University, Boston, MA 02215.
George G
GG
Daaboul
Electrical & Computer Engineering Dept., Boston University, Boston, MA 02215.
Carlos A
CA
Lopez
Electrical & Computer Engineering Dept., Boston University, Boston, MA 02215.
Jyothsna
J
Chinnala
Electrical & Computer Engineering Dept., Boston University, Boston, MA 02215.|Biomedical Engineering Dept., Boston University, Boston, MA 02215.|Physics Department, Boston University, Boston, MA 02215.
Bennett B
BB
Goldberg
Department of Microbiology and National Emerging and Infectious Diseases Laboratories, Boston University School of Medicine, Boston, MA 02118.|Boston University Photonics Center, Boston University, Boston, MA 02215.
John H
JH
Connor
Electrical & Computer Engineering Dept., Boston University, Boston, MA 02215.|Biomedical Engineering Dept., Boston University, Boston, MA 02215.|Physics Department, Boston University, Boston, MA 02215.
M Selim
MS
Ünlü
D000818:Animals; D015374:Biosensing Techniques; D018076:DNA, Complementary / Q000378:metabolism; D004279:DNA, Viral / Q000032:analysis; D006023:Glycoproteins / Q000737:chemistry; D019142:Hemorrhagic Fever, Ebola / Q000175:diagnosis* / Q000821:virology*; D006801:Humans; D007368:Interferometry; D008024:Ligands; D057230:Limit of Detection; D008379:Marburg Virus Disease / Q000175:diagnosis* / Q000821:virology*; D053758:Nanoparticles / Q000737:chemistry; D036103:Nanotechnology / Q000379:methods; D016011:Normal Distribution; D020411:Oligonucleotide Array Sequence Analysis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D018116:Vesiculovirus
D018076:DNA, Complementary; D004279:DNA, Viral; D006023:Glycoproteins; D008024:Ligands