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Technology Description	We describe a new approach for infectious disease surveillance that facilitates rapid identification of known and emerging pathogens. The process uses broad-range polymerase chain reaction (PCR) to amplify nucleic acid targets from large groupings of organisms, electrospray ionization mass spectrometry for accurate mass measurements of PCR products, and base composition signature analysis to identify organisms in a sample. We demonstrate this principle by using 14 isolates of 9 diverse Coronavirus spp., including the severe acute respiratory syndrome–associated coronavirus (SARS-CoV).
Category	Technology
PRC	Scripps Research Institute
PubMed ID	15757550
Author	Rangarajan Sampath, Steven A. Hofstadler, Lawrence B. Blyn, Mark W. Eshoo, Thomas A. Hall, Christian Massire, Harold M. Levene, James C. Hannis, Patina M. Harrell, Benjamin Neuman, Michael J. Buchmeier, Yun Jiang, Raymond Ranken, Jared J. Drader, Vivek Samant, Richard H. Griffey, John A. McNeil, Stanley T. Crooke, and David J. Ecker
Publication Description	We present an alternative approach for rapid, sensitive, and high-throughput detection of infectious organisms. We use broad-range PCR to generate amplification products from the broadest possible grouping of organisms, followed by electrospray ionization mass spectrometry and base composition analysis of the products.
Methodology	The base compositions of strategically selected regions of the genome are used to identify and distinguish organisms in the sample. Enhanced breadth of priming is achieved through the use of primers and probes containing 5-propynyl deoxycytidine and deoxyuridine nucleotides that offer increased affinity and base pairing selectivity. Positioning the 5-propynyl primidine-modified nucleotides at highly conserved positions enables priming at short consensus regions and significantly increases the extent to which broad groups of organisms can be amplified.

National Institute of Allergy and Infectious Disease