A qPCR Assay for the Quantification of Selected Genotypic Variants of Spodoptera frugiperda Multiple Nucleopolyhedrovirus ( Baculoviridae )

Alphabaculoviruses are lethal dsDNA viruses of Lepidoptera that have high genetic diversity and are transmitted in aggregates within proteinaceous occlusion bodies. This mode of transmission has implications for their efficacy as biological insecticides. A Nicaraguan isolate of Spodoptera frugiperda...

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Bibliographic Details
Published in:Viruses 2024-06, Vol.16 (6), p.881
Main Authors: Molina-Ruiz, Cindy S, Zamora-Briseño, Jesús Alejandro, Simón, Oihane, Lasa, Rodrigo, Williams, Trevor
Format: Article
Language:English
Subjects:
alphabaculovirus
Animals
Baculoviridae
Baculoviruses
Cloning
copy number determination
Design
Disease transmission
Diseases
DNA, Viral - genetics
efficiency
fall armyworm
Genes
Genetic aspects
Genetic diversity
Genetic Variation
Genomes
Genotype
Genotypes
genotypic diversity
Health aspects
Host-virus relationships
Infections
Laboratories
Lepidoptera
Nucleopolyhedroviruses - classification
Nucleopolyhedroviruses - genetics
Nucleopolyhedroviruses - isolation & purification
Occlusion bodies
Plasmids
Polyhedrin
Real-Time Polymerase Chain Reaction - methods
Spodoptera - virology
Spodoptera frugiperda
Virions
Viruses
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Summary:Alphabaculoviruses are lethal dsDNA viruses of Lepidoptera that have high genetic diversity and are transmitted in aggregates within proteinaceous occlusion bodies. This mode of transmission has implications for their efficacy as biological insecticides. A Nicaraguan isolate of Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV-NIC) comprising nine genotypic variants has been the subject of considerable study due to the influence of variant interactions on the insecticidal properties of mixed-variant occlusion bodies. As part of a systematic study on the replication and transmission of variant mixtures, a tool for the accurate quantification of a selection of genotypic variants was developed based on the quantitative PCR technique (qPCR). First, primer pairs were designed around a region of high variability in four variants named SfNic-A, SfNic-B, SfNic-C and SfNic-E to produce amplicons of 103-150 bp. Then, using cloned purified amplicons as standards, amplification was demonstrated over a dynamic range of 10 -10 copies of each target. The assay was efficient (mean ± SD: 98.5 ± 0.8%), reproducible, as shown by low inter- and intra-assay coefficients of variation (
ISSN:1999-4915
1999-4915
DOI:10.3390/v16060881