Metagenomic arbovirus detection using MinION nanopore sequencing

被引:49
作者
Batovska, Jana [1 ,2 ]
Lynch, Stacey E. [1 ]
Rodoni, Brendan C. [1 ,2 ]
Sawbridge, Tim I. [1 ,2 ]
Cogan, Noel O. I. [1 ,2 ]
机构
[1] Agr Victoria Res, AgriBio Ctr AgriBiosci, 5 Ring Rd, Bundoora, Vic 3083, Australia
[2] La Trobe Univ, Sch Appl Syst Biol, Bundoora, Vic 3086, Australia
关键词
Mosquito; Culicidae; Ross River virus; MinION; MiSeq; Surveillance; REAL-TIME; IDENTIFICATION; BACTERIAL; VIRUS; ZIKA;
D O I
10.1016/j.jviromet.2017.08.019
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
With its small size and low cost, the hand-held MinION sequencer is a powerful tool for in-field surveillance. Using a metagenomic approach, it allows non-targeted detection of viruses in a sample within a few hours. This study aimed to determine the ability of the MinION to metagenomically detect and characterise a virus from an infected mosquito. RNA was extracted from an Aedes notoscriptus mosquito infected with Ross River virus (RRV), converted into cDNA and sequenced on the MinION. Bioinformatic analysis of the MinION reads led to detection of full-length RRV, with reads of up to 2.5 kb contributing to the assembly. The cDNA was also sequenced on the MiSeq sequencer, and both platforms recovered the RRV genome with > 98% accuracy. This proof of concept study demonstrates the metagenomic detection of an arbovirus, using the MinION, directly from a mosquito with minimal sample purification.
引用
收藏
页码:79 / 84
页数:6
相关论文
共 37 条
[1]  
[Anonymous], SCI REP UK
[2]   Applications of Next-Generation Sequencing Technologies to Diagnostic Virology [J].
Barzon, Luisa ;
Lavezzo, Enrico ;
Militello, Valentina ;
Toppo, Stefano ;
Palu, Giorgio .
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 2011, 12 (11) :7861-7884
[3]  
Batovska J., 2017, MOL ECOL RESOUR
[4]   Molecular identification of mosquitoes (Diptera: Culicidae) in southeastern Australia [J].
Batovska, Jana ;
Blacket, Mark J. ;
Brown, Karen ;
Lynch, Stacey E. .
ECOLOGY AND EVOLUTION, 2016, 6 (09) :3001-3011
[5]   Nanocall: an open source basecaller for Oxford Nanopore sequencing data [J].
David, Matei ;
Dursi, L. J. ;
Yao, Delia ;
Boutros, Paul C. ;
Simpson, Jared T. .
BIOINFORMATICS, 2017, 33 (01) :49-55
[6]   Enrichment by hybridisation of long DNA fragments for Nanopore sequencing [J].
Eckert, Sabine E. ;
Chan, Jackie Z. -M. ;
Houniet, Darren ;
Breuer, Judy ;
Speight, Graham .
MICROBIAL GENOMICS, 2016, 2 (09) :e000087
[7]  
Garalde D. R., 2016, 068809 BIORXIV
[8]   Rapid metagenomic identification of viral pathogens in clinical samples by real-time nanopore sequencing analysis [J].
Greninger, Alexander L. ;
Naccache, Samia N. ;
Federman, Scot ;
Yu, Guixia ;
Mbala, Placide ;
Bres, Vanessa ;
Stryke, Doug ;
Bouquet, Jerome ;
Somasekar, Sneha ;
Linnen, Jeffrey M. ;
Dodd, Roger ;
Mulembakani, Prime ;
Schneider, Bradley S. ;
Muyembe-Tamfum, Jean-Jacques ;
Stramer, Susan L. ;
Chiu, Charles Y. .
GENOME MEDICINE, 2015, 7
[9]  
Hall R., 2011, MOL DETECTION HUMAN, P349
[10]   Nanopore Sequencing as a Rapidly Deployable Ebola Outbreak Tool [J].
Hoenen, Thomas ;
Groseth, Allison ;
Rosenke, Kyle ;
Fischer, Robert J. ;
Hoenen, Andreas ;
Judson, Seth D. ;
Martellaro, Cynthia ;
Falzarano, Darryl ;
Marzi, Andrea ;
Squires, R. Burke ;
Wollenberg, Kurt R. ;
de Wit, Emmie ;
Prescott, Joseph ;
Safronetz, David ;
van Doremalen, Neeltje ;
Bushmaker, Trenton ;
Feldmann, Friederike ;
McNally, Kristin ;
Bolay, Fatorma K. ;
Fields, Barry ;
Sealy, Tara ;
Rayfield, Mark ;
Nichol, Stuart T. ;
Zoon, Kathryn C. ;
Massaquoi, Moses ;
Munster, Vincent J. ;
Feldmann, Heinz .
EMERGING INFECTIOUS DISEASES, 2016, 22 (02) :331-334