Rapid detection and molecular serotyping of adenovirus by use of PCR followed by electrospray ionization mass spectrometry.
We have developed a PCR/electrospray ionization mass spectrometry (PCR/ESI-MS) assay for the rapid detection, identification, and serotyping of human adenoviruses. The assay employs a high-performance mass spectrometer to "weigh" the amplicons obtained from PCR using primers designed to amplify known human adenoviruses. Masses are converted to base compositions and, by comparison against a database of the genetic sequences, the serotype present in a sample is determined. The performance of the assay was demonstrated with quantified viral standards and environmental and human clinical samples collected from a military training facility. Over 500 samples per day can be analyzed with sensitivities greater than 100 genomes per reaction. This approach can be applied to many other families of infectious agents for rapid and sensitive analysis.
Blyn
Lawrence B
LB
Ibis Biosciences, Inc., Carlsbad, CA 92008, USA. lblyn@isisph.com
Hall
Thomas A
TA
Libby
Brian
B
Ranken
Raymond
R
Sampath
Rangarajan
R
Rudnick
Karl
K
Moradi
Emily
E
Desai
Anjali
A
Metzgar
David
D
Russell
Kevin L
KL
Freed
Nikki E
NE
Balansay
Melinda
M
Broderick
Michael P
MP
Osuna
Miguel A
MA
Hofstadler
Steven A
SA
Ecker
David J
DJ
D000256:Adenoviridae / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D000257:Adenoviridae Infections / Q000175:diagnosis* / Q000821:virology*; D002695:Chlamydiales; D017931:DNA Primers / Q000235:genetics; D001330:Electronic Data Processing; D004783:Environmental Microbiology; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D012703:Serotyping / Q000379:methods; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D017931:DNA Primers