Detection and identification of baculovirus pesticides by multitemperature single-strand conformational polymorphism.
The method of single-strand conformational polymorphism (SSCP) was modified in our laboratories for the characterization of baculoviruses, insect viruses with great potential for use as bioinsecticides in biological protection programs. A series of primers were synthesized after the comparison of the polyhedrin gene sequences of over 20 baculoviruses. Polyhedrin is a highly conserved protein which is responsible for the persistence of the virus in the environment. Universal primers were designed which could be used in polymerase chain reactions (PCR) containing genomic DNA from an array of nucleopolyhedrosis viruses (NPVs) including these which are used as biopesticides against important pests of forests and crops, such as Anticarsia gemmatalis, Spodoptera frugiperda, Lymantria dispar, Lymantria monacha and many others. PCR products were denatured and subjected to single-strand DNA electrophoresis at variable temperatures (MSSCP) where, after silver staining, they gave ssDNA band patterns characteristic for each baculovirus species. This technique can be potentially applied to detect baculoviruses in insects collected in the field, as well as to plant tissues and the excrements or bodies of predators without need for sequencing the PCR products. Sometimes MSSCP can be used not only for species determination but also as an indication of genomic variability which can be related to infectivity.
Szewczyk
Boguslaw
B
Department of Molecular Virology, Intercollegiate Faculty of Biotechnology of the University of Gdansk and Medical University of Gdansk, Gdansk, Poland. szewczyk@biotech.ug.gda.pl
Barski
Piotr
P
Sihler
William
W
Rabalski
Lukasz
L
Skrzecz
Iwona
I
Hoyos-Carvajal
Liliana
L
de Souza
Marlinda Lobo
ML
D000818:Animals; D016367:Baculoviridae / Q000145:classification* / Q000235:genetics*; D014644:Genetic Variation; D007313:Insecta / Q000821:virology; D010572:Pest Control, Biological; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational / Q000235:genetics*; D013045:Species Specificity; D015678:Viral Structural Proteins / Q000235:genetics
D015678:Viral Structural Proteins