New method for differentiation of granuloviruses (Betabaculoviruses) based on real-time polymerase chain reaction (real-time PCR)

Martyna Krejmer-Rąbalska , Łukasz Rąbalski , Michael D. Jukes , Marlinda Lobo de Souza , Sean D. Moore , Bogusław Szewczyk

Abstract

Baculoviridae is a highly diverse family of rod-shaped viruses with double-stranded DNA. To date, almost 100 species have had their complete genomic sequences deposited in the GenBank database, a quarter of which comprises granuloviruses (GVs). Many of the genomes are sequenced using next-generation sequencing, which is currently considered the best method for characterizing new species, but it is time-consuming and expensive. Baculoviruses form a safe alternative to overused chemical pesticides and therefore there is a constant need for identifying new species that can be active components of novel biological insecticides. In this study, we have described a fast and reliable method for the detection of new and differentiation of previously analyzed granulovirus species based on a real-time polymerase chain reaction (PCR) technique with melting point curve analysis. The sequences of highly conserved baculovirus genes, such as granulin and late expression factors 8 and 9 (lef-8 and lef-9), derived from GVs available to date have been analyzed and used for degenerate primer design. The developed method was tested on a representative group of eight betabaculoviruses with comparisons of melting temperatures to allow for quick and preliminary granulovirus detection. The proposed real-time PCR procedure may be a very useful tool as an easily accessible screening method in a majority of laboratories.
Author Martyna Krejmer-Rąbalska (IFB)
Martyna Krejmer-Rąbalska,,
- Intercollegiate Faculty of Biotechnology UG
, Łukasz Rąbalski (IFB / IB / LRV)
Łukasz Rąbalski,,
- Laboratory of Recombinant Vaccines
, Michael D. Jukes
Michael D. Jukes,,
-
, Marlinda Lobo de Souza
Marlinda Lobo de Souza,,
-
, Sean D. Moore
Sean D. Moore,,
-
, Bogusław Szewczyk (IFB / IB / LRV)
Bogusław Szewczyk,,
- Laboratory of Recombinant Vaccines
Journal seriesViruses-Basel, ISSN 1999-4915, (A 30 pkt)
Issue year2019
Vol11
No2
Pages1-16
Publication size in sheets0.75
Article number115
Keywords in Englishbetabaculovirus, detection, real-time PCR, lef-9, lef-8, granulin
ASJC Classification2406 Virology; 2725 Infectious Diseases
DOIDOI:10.3390/v11020115
URL https://doi.org/10.3390/v11020115
Languageen angielski
LicenseJournal (articles only); published final; Uznanie Autorstwa (CC-BY); with publication
Score (nominal)30
ScoreMinisterial score = 30.0, 24-07-2019, ArticleFromJournal
Publication indicators Scopus SNIP (Source Normalised Impact per Paper): 2016 = 1.018; WoS Impact Factor: 2017 = 3.761 (2) - 2017=3.737 (5)
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