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
AbstractBaculoviridae 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.
|Journal series||Viruses-Basel, ISSN 1999-4915, (N/A 100 pkt)|
|Publication size in sheets||0.75|
|Keywords in English||betabaculovirus, detection, real-time PCR, lef-9, lef-8, granulin|
|License||Journal (articles only); published final; ; with publication|
|Score||= 100.0, 17-11-2019, ArticleFromJournal|
|Publication indicators||: 2016 = 1.018; : 2018 = 3.811 (2) - 2018=3.916 (5)|
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