Artikel
SARS-CoV transmission epidemiology revealed by MALDI-TOF mass spectrometry-based viral genotyping
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Veröffentlicht: | 26. Mai 2004 |
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Gliederung
Text
Epidemiological investigations of infectious disease are mainly dependent on indirect contact information and only occasionally assisted by viral sequence variation characterization from clinical isolates. Direct sequence analysis of virus, particularly at a population level, is generally thought to be too cumbersome, technically difficult and expensive. We present here a matrix-assisted laser desorption ionization/time-of-flight (MALDI-TOF) MS-based high throughput genotyping approach that overcomes these problems and apply it to the severe acute respiratory syndrome (SARS) outbreak in Singapore. We first determined the high sensitivity of the MS genotyping analysis for detecting SARS-coronavirus (CoV) genotypes by analyzing human blood samples spiked with SARS-CoV, and then genotyped 13 Singapore SARS-CoV isolates on 18 single nucleotide variations (SNVs). Our results identified 9 SNV positions whose genotypes could define the molecular relationship between the Singapore SARS-CoV isolates and further pointed to a new, previously unidentified, primary route of introduction of SARS-CoV into the Singapore population. In a separate analysis we determined the virus genotype directly from a clinical sample to clarify an unresolved epidemiological link regarding the acquisition of SARS in a German patient. Our study demonstrated that the MALDI-TOF MS-based genotyping analysis is a sensitive and accurate platform for characterizing SARS-CoV genetic variations in clinical samples and will allow a high throughput approach to epidemiological investigations which will greatly facilitate their accuracy.