Detection and kinetic analysis of Epinotia aporema granulovirus in its lepidopteran host by real-time PCR

Abstract

Epinotia aporema granulovirus (EpapGV) has attracted interest as a potential biocontrol agent of the soybean pest Epinotia aporema in Argentina. Studies on virus/host interactions conducted so far have lacked an accurate method to assess the progress of virus load during the infection process. The present paper reports the development of a real-time PCR for EpapGV and its application to describe viral kinetics following ingestion of two different virus doses by last-instar E. aporema larvae. Real-time PCR was shown to be a reliable method to detect and quantify the presence of EpapGV in the analyzed samples. The increase in virus titer (log) exhibited a sigmoidal pattern, with an exponential growth phase between 24 and 48 h postinfection for both initial doses tested.

Contribution of a parasitoid species to multiplication and transmission of a multiple nucleopolyhedrovirus in caterpillars

“…Caballero, 2006). Recently, the emergence of RT-qPCR has improved our ability to accurately quantify insect viruses (Arneodo et al, 2012). In this study, we manipulated these methods to demonstrate the interactions of S. exigua-SeMNPV-M. pallidipes.…”

Degenerate Primed Polymerase Chain Reaction for Detection of Both Nucleopolyhedrovirus and Granulovirus

“…Moreover, serological techniques are time consuming, require highly experienced personnel and are less precise than nucleotide sequence determination. Generally, the use of PCR technology for virus detection, identification and characterization is a basic tool in many virology laboratories (Moraes and Maruniak, 1997, Moraes et al, 1999, Wang et al, 2000, Christian et al, 2001, Moraes and Maruniak, 2001, Woo, 2001, Ikuno et al, 2004, Lange et al, 2004, Jehle et al, 2006, Murillo et al, 2006, Kundu et al, 2008, Manzán et al, 2008, Galal, 2009, Hewson et al, 2011, Ravikumar et al, 2011, Arneodo et al, 2012. Indeed, a good set of primers for simultaneous NPV and GV detection is a powerful tool for large Baculovirus sample screening.…”

“…Recently, Arneodo et al (2012) developed a realtime PCR approach for detection and and kinetic analysis of Epinotia aporema granulovirus in its host.…”

“…In addition, the commercialization and release of recombinant viruses in the environment 2 created the concern that they might cause ecological disturbances, such as displacement of native microorganisms, adverse effects on non-target organisms and the horizontal transfer of DNA into non-target organisms (Leung et al, 1994). For the above mentioned reasons, many authors were interested in developing an accurate and easy diagnostic method for early, reliable and rapid detection of NPV and GV infections (Wang et al, 2000, Christian et al, 2001, Moraes and Maruniak, 2001, Woo, 2001, Ikuno et al, 2004, Lange et al, 2004, Jehle et al, 2006, Murillo et al, 2006, Kundu et al, 2008, Manzán et al, 2008, Galal, 2009, Hewson et al, 2011, Ravikumar et al, 2011, Arneodo et al, 2012. Several methods have been employed to detect wild type or recombinant baculoviruses, such as microscopic diagnosis (Traverner and Connor, 1992), serological techniques (Brown et al, 1982, Naser and Miltenburger, 1983, Webb and Shelton, 1990, radioimmunoassay techniques (Smith andSummers, 1981, Knell et al, 1983), and DNA dot blot hybridization assays (Ward et al, 1987, Keating et al, 1989.…”

“…PCR has been extensively used to detect many organisms such as animal, human, plant and various pathogens. Many authors reported the use of PCR technology to screen baculoviruses in different approaches over the last three decades (Webb et al, 1991, Burand et al, 1992, Kundu et al, 2003, Ikuno et al, 2004, Kundu et al, 2008, Galal, 2009, Manzán et al, 2008, Hewson et al, 2011, Ravikumar et al, 2011, Arneodo et al, 2012.…”