Genome reconstruction of white spot syndrome virus (WSSV) from archival Davidson's-fixed paraffin embedded shrimp (Penaeus vannamei) tissue

被引:15
作者
Cruz-Flores, Roberto [1 ]
Mai, Hung N. [1 ]
Kanrar, Siddhartha [1 ]
Caro, Luis Fernando Aranguren [1 ]
Dhar, Arun K. [1 ]
机构
[1] Univ Arizona, Aquaculture Pathol Lab, Sch Anim & Comparat Biomed Sci, Bldg 90, Tucson, AZ 85721 USA
关键词
SEQUENCE; SAMPLES; DNA;
D O I
10.1038/s41598-020-70435-x
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Formalin-fixed paraffin-embedded (FFPE) tissues are a priceless resource for diagnostic laboratories worldwide. However, DNA extracted from these tissues is often not optimal for most downstream molecular analysis due to fragmentation and chemical modification. In this study, the complete genome of white spot syndrome virus (WSSV) was reconstructed from similar to 2-year-old archived Davidson's-fixed paraffin-embedded (DFPE) shrimp tissue using Next Generation Sequencing (NGS). A histological analysis was performed on archived DFPE shrimp tissue and a sample showing a high level of WSSV infection was selected for molecular analysis. The viral infection was further confirmed by molecular methods. DNA isolated from DFPE and fresh frozen (FF) tissues were sequenced by NGS. The complete genome reconstruction of WSSV (similar to 305 kbp) was achieved from both DFPE and FF tissue. Single nucleotide polymorphisms, insertion and deletions were compared between the genomes. Thirty-eight mutations were identified in the WSSV genomes from the DFPE and FF that differed from the reference genome. This is the first study that has successfully sequenced the complete genome of a virus of over 300 kbp from archival DFPE tissue. These findings demonstrate that DFPE shrimp tissue represents an invaluable resource for prospective and retrospective studies, evolutionary studies and opens avenues for pathogen discovery.
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页数:10
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共 34 条
[1]   Evaluation of the preservation of shrimp samples with Davidson's AFA fixative for infectious myonecrosis virus (IMNV) in situ hybridization [J].
Andrade, Thales P. D. ;
Redman, Rita M. ;
Lightner, Donald V. .
AQUACULTURE, 2008, 278 (1-4) :179-183
[2]   The RAST server: Rapid annotations using subsystems technology [J].
Aziz, Ramy K. ;
Bartels, Daniela ;
Best, Aaron A. ;
DeJongh, Matthew ;
Disz, Terrence ;
Edwards, Robert A. ;
Formsma, Kevin ;
Gerdes, Svetlana ;
Glass, Elizabeth M. ;
Kubal, Michael ;
Meyer, Folker ;
Olsen, Gary J. ;
Olson, Robert ;
Osterman, Andrei L. ;
Overbeek, Ross A. ;
McNeil, Leslie K. ;
Paarmann, Daniel ;
Paczian, Tobias ;
Parrello, Bruce ;
Pusch, Gordon D. ;
Reich, Claudia ;
Stevens, Rick ;
Vassieva, Olga ;
Vonstein, Veronika ;
Wilke, Andreas ;
Zagnitko, Olga .
BMC GENOMICS, 2008, 9 (1)
[3]   Virus characterization and discovery in formalin-fixed paraffin-embedded tissues [J].
Bodewes, Rogier ;
van Run, Peter R. W. A. ;
Schurch, Anita C. ;
Koopmans, Marion P. G. ;
Osterhaus, Albert D. M. E. ;
Baumgaertner, Wolfgang ;
Kuiken, Thijs ;
Smits, Saskia L. .
JOURNAL OF VIROLOGICAL METHODS, 2015, 214 :54-59
[4]   Detection of viral pathogens in high grade gliomas from unmapped next-generation sequencing data [J].
Cimino, Patrick J. ;
Zhao, Guoyan ;
Wang, David ;
Sehn, Jennifer K. ;
Lewis, James S., Jr. ;
Duncavage, Eric J. .
EXPERIMENTAL AND MOLECULAR PATHOLOGY, 2014, 96 (03) :310-315
[5]   ENZOOTIC NUCLEAR POLYHEDROSIS-VIRUS OF PINK SHRIMP - ULTRASTRUCTURE, PREVALENCE, AND ENHANCEMENT [J].
COUCH, JA .
JOURNAL OF INVERTEBRATE PATHOLOGY, 1974, 24 (03) :311-331
[6]   Mauve: Multiple alignment of conserved genomic sequence with rearrangements [J].
Darling, ACE ;
Mau, B ;
Blattner, FR ;
Perna, NT .
GENOME RESEARCH, 2004, 14 (07) :1394-1403
[7]   Diversity of single-stranded DNA containing viruses in shrimp [J].
Dhar A.K. ;
Cruz-Flores R. ;
Caro L.F.A. ;
Siewiora H.M. ;
Jory D. .
VirusDisease, 2019, 30 (1) :43-57
[8]   Characterization of a Taura syndrome virus isolate originating from the 2004 Texas epizootic in cultured shrimp [J].
Dhar, Arun K. ;
Lakshman, Dilip K. ;
Amundsen, Keenan ;
Robles-Sikisaka, Refugio ;
Kaizer, Krista N. ;
Roy, Sribash ;
Hasson, Kenneth W. ;
Allnutt, F. C. Thomas .
ARCHIVES OF VIROLOGY, 2010, 155 (03) :315-327
[9]  
Dupinay T, 2012, CURR TOP VIROL, V10, P64
[10]   Quantitative real time PCR for the measurement of white spot syndrome virus in shrimp [J].
Durand, SV ;
Lightner, DV .
JOURNAL OF FISH DISEASES, 2002, 25 (07) :381-389