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Rapid identification of emerging pathogens: coronavirus
Authors:Sampath Rangarajan  Hofstadler Steven A  Blyn Lawrence B  Eshoo Mark W  Hall Thomas A  Massire Christian  Levene Harold M  Hannis James C  Harrell Patina M  Neuman Benjamin  Buchmeier Michael J  Jiang Yun  Ranken Raymond  Drader Jared J  Samant Vivek  Griffey Richard H  McNeil John A  Crooke Stanley T  Ecker David J
Affiliation:Ibis Therapeutics, 1891 Rutherford Ct, Carlsbad, CA 92008, USA. rsampath@isisph.com
Abstract: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). We show that this method could identify and distinguish between SARS and other known CoV, including the human CoV 229E and OC43, individually and in a mixture of all 3 human viruses. The sensitivity of detection, measured by using titered SARS-CoV spiked into human serum, was approximate, equals1 PFU/mL. This approach, applicable to the surveillance of bacterial, viral, fungal, or protozoal pathogens, is capable of automated analysis of >900 PCR reactions per day.
Keywords:PCR   molecular epidemiology   emerging pathogens   SARS virus   infectious disease surveillance   ESI mass spectrometry   research
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