Identification of viruses using microfluidic protein profiling and Bayesian classification.
We present a rapid method for the identification of viruses using microfluidic chip gel electrophoresis (CGE) of high-copy number proteins to generate unique protein profiles. Viral proteins are solubilized by heating at 95 degrees C in borate buffer containing detergent (5 min), then labeled with fluorescamine dye (10 s), and analyzed using the microChemLab CGE system (5 min). Analyses of closely related T2 and T4 bacteriophage demonstrate sufficient assay sensitivity and peak resolution to distinguish the two phage. CGE analyses of four additional viruses--MS2 bacteriophage, Epstein-Barr, respiratory syncytial, and vaccinia viruses--demonstrate reproducible and visually distinct protein profiles. To evaluate the suitability of the method for unique identification of viruses, we employed a Bayesian classification approach. Using a subset of 126 replicate electropherograms of the six viruses and phage for training purposes, successful classification with non-training data was 66/69 or 95% with no false positives. The classification method is based on a single attribute (elution time), although other attributes such as peak width, peak amplitude, or peak shape could be incorporated and may improve performance further. The encouraging results suggest a rapid and simple way to identify viruses without requiring specialty reagents such as PCR probes and antibodies.
Fruetel
Julia A
JA
Sandia National Laboratories, Livermore, California 94551-0969, USA. jfruet@sandia.gov
West
Jason A A
JA
Debusschere
Bert J
BJ
Hukari
Kyle
K
Lane
Todd W
TW
Najm
Habib N
HN
Ortega
Jose
J
Renzi
Ronald F
RF
Shokair
Isaac
I
VanderNoot
Victoria A
VA
D001435:Bacteriophages / Q000737:chemistry; D002138:Calibration; D046209:Electrophoresis, Microchip / Q000191:economics / Q000295:instrumentation* / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004867:Equipment Design; D046210:Microfluidic Analytical Techniques / Q000191:economics / Q000295:instrumentation* / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors; D014764:Viral Proteins / Q000032:analysis*; D014780:Viruses / Q000737:chemistry*
D014764:Viral Proteins