Open AccessRapid Detection and Molecular Serotyping of Adenovirus by Use of PCR Followed by Electrospray Ionization Mass Spectrometry
Author(s) -
Lawrence B. Blyn,
Thomas A. Hall,
Brian Libby,
Raymond Ranken,
Rangarajan Sampath,
Karl Rudnick,
Emily Moradi,
Anjali Desai,
David Metzgar,
Kevin L. Russell,
Nikki E. Freed,
Melinda Balansay,
Michael Broderick,
Miguel Osuna,
Steven A. Hofstadler,
David J. Ecker
Publication year - 2008
Publication title -
journal of clinical microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.349
H-Index - 255
eISSN - 1070-633X
pISSN - 0095-1137
DOI - 10.1128/jcm.00801-07
Subject(s) - amplicon , serotype , mass spectrometry , electrospray ionization , chromatography , polymerase chain reaction , biology , electrospray , virology , microbiology and biotechnology , chemistry , genetics , gene
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.