Mass Spectrometry-Based Proteolytic Mapping for Rapid Virus Identification

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• 作者：Zhong-Ping Yao ; Plamen A. Demirev ; and Catherine Fenselau
• 刊名：Analytical Chemistry
• 出版年：2002
• 出版时间：June 1, 2002
• 年：2002
• 卷：74
• 期：11
• 页码：2529 - 2534
• 全文大小：67K
• 年卷期：v.74,no.11(June 1, 2002)
• ISSN：1520-6882

文摘

A novel method is proposed for rapid identification ofviruses and other organisms that show a low number ofbiomarkers, based on the construction of databases oforganism-specific tryptic peptide masses. The peptideproducts of any protease that cuts at specific residues canbe accommodated. Experimentally, a sample of intactvirus, e.g., one collected from the atmosphere, is digestedwith a selective protease for a short time, and the digestionproducts are analyzed by MALDI-TOF mass spectrometrywithout fractionation or purification. In the present proofof concept, the Sindbis virus AR 339 was identified byusing the masses of observed tryptic peptide products toquery a database composed of tryptic peptide massesgenerated in silico for six viruses whose genomes havebeen sequenced. Two algorithms were tested for identification-a direct score-ranking algorithm and an algorithm that evaluates the probability of random matching.The Sindbis virus was unambiguously identified by eitherapproach. The influence of factors such as experimentalmass accuracy, number of missed cleavages, and database size on the identification algorithms has also beenevaluated, with the objective of extending the approachto other microorganisms.