Author: Barzon, Luisa; Lavezzo, Enrico; Militello, Valentina; Toppo, Stefano; Palù, Giorgio
Title: Applications of Next-Generation Sequencing Technologies to Diagnostic Virology Document date: 2011_11_14
ID: 01nuj0lk_9
Snippet: One of the first applications of NGS for pathogen discovery was the investigation of three patients who died of a febrile illness a few weeks after transplantation of solid organs from a single donor and for whom conventional microbiological and molecular tests, as well as microarray analysis for a wide range of infectious agents, had not been informative [21] . In this study, RNA was purified from blood, cerebrospinal fluid and tissue specimens .....
Document: One of the first applications of NGS for pathogen discovery was the investigation of three patients who died of a febrile illness a few weeks after transplantation of solid organs from a single donor and for whom conventional microbiological and molecular tests, as well as microarray analysis for a wide range of infectious agents, had not been informative [21] . In this study, RNA was purified from blood, cerebrospinal fluid and tissue specimens from transplant recipients and, after digestion with DNase to eliminate human DNA, RNA was reverse-transcribed and amplified with random primers. Amplification products were pooled and sequenced with the use of the 454 pyrosequencing platform. After subtraction of sequences of vertebrates and highly repetitive sequences, contiguous sequences were assembled and compared with motifs represented in databases of microbes, leading to the identification of putative protein sequences which were consistent with an Old World arenavirus.
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