scholarly journals Use of Sequence-Independent, Single-Primer-Amplification (SISPA) for rapid detection, identification, and characterization of avian RNA viruses

Virology ◽  
2017 ◽  
Vol 509 ◽  
pp. 159-166 ◽  
Author(s):  
Klaudia Chrzastek ◽  
Dong-hun Lee ◽  
Diane Smith ◽  
Poonam Sharma ◽  
David L. Suarez ◽  
...  
Author(s):  
Noam Auslander ◽  
Ayal B. Gussow ◽  
Sean Benler ◽  
Yuri I. Wolf ◽  
Eugene V. Koonin

SummaryAdvances in metagenomics enable massive discovery of diverse, distinct microbes and viruses. Bacteriophages, the most abundant biological entity on Earth, evolve rapidly, and therefore, detection of unknown bacteriophages in sequence datasets is a challenge. The existing methods rely on sequence similarity to known bacteriophage sequences, impeding the identification and characterization of distinct bacteriophage families. We present Seeker, a deep-learning tool for reference-free identification of phage sequences. Seeker allows rapid detection of phages in sequence datasets and clean differentiation of phage sequences from bacterial ones, even for phages with little sequence similarity to established phage families. We comprehensively validate Seeker’s ability to identify unknown phages and employ Seeker to detect unknown phages, some of which are highly divergent from known phage families. We provide a web portal (seeker.pythonanywhere.com) and a user-friendly python package (https://github.com/gussow/seeker) allowing researchers to easily apply Seeker in metagenomic studies, for the detection of diverse unknown bacteriophages.


2016 ◽  
Vol 22 (8) ◽  
pp. 1448-1451 ◽  
Author(s):  
Andy Kilianski ◽  
Pierce A. Roth ◽  
Alvin T. Liem ◽  
Jessica M. Hill ◽  
Kristen L. Willis ◽  
...  

2011 ◽  
Vol 74 (1) ◽  
pp. 13-17 ◽  
Author(s):  
V. LORUSSO ◽  
A. DAMBROSIO ◽  
N. C. QUAGLIA ◽  
A. PARISI ◽  
G. LASALANDRA ◽  
...  

Verocytotoxin-producing Escherichia coli (VTEC) O26 is an emergent pathotype that has caused an increasing number of sporadic cases and outbreaks of gastroenteritis, hemorrhagic colitis, and hemolytic uremic syndrome in the United States and Europe. Many cases are associated with the consumption of milk and undercooked or fermented meats. The stx2 strains of VTEC O26 seem to be more likely to cause human infections than isolates expressing only stx1. The isolation and identification of VTEC O26 from foods is labor intensive and time-consuming. We developed a multiplex PCR (M-PCR) assay for the identification and characterization of E. coli O26 VTEC and its detection in raw milk and ground beef. The method is based on the amplification of the wzx, stx1, and stx2 genes for the simultaneous detection of the O26 antigen and verocytotoxin types 1 and 2. This M-PCR assay had a sensitivity of 108 CFU/ml when applied to a bacterial suspension and of 106 CFU/ml or g when applied to both inoculated milk and minced beef samples. This M-PCR assay also was highly specific, and results were consistently negative for negative controls (nonpathogenic E. coli strains, uninoculated milk and beef samples, and samples inoculated with the nontarget microorganisms). This method could be used for the rapid detection of E. coli O26 VTEC from foods and for the rapid identification and characterization of clinical and environmental isolates.


PLoS ONE ◽  
2016 ◽  
Vol 11 (5) ◽  
pp. e0153881 ◽  
Author(s):  
Guillaume Carissimo ◽  
Karin Eiglmeier ◽  
Julie Reveillaud ◽  
Inge Holm ◽  
Mawlouth Diallo ◽  
...  

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