Production and Application of Polyclonal Antibodies Against Recombinant Capsid Protein of Extra Small Virus of Macrobrachium rosenbergii

Abstract

Macrobrachium rosenbergii nodavirus along with a satellite virus, extra small virus (XSV) causes white tail disease (WTD) in the giant freshwater prawn M. rosenbergii. Infected M. rosenbergii postlarvae were collected from a hatchery in Kakinada, Andhra Pradesh. The gene coding the capsid protein of XSV was cloned in a bacterial expression vector pRSET A and the recombinant protein was expressed in Escherichia coli BL21(DE3)pLysS cells. The recombinant protein was purified by Nickel affinity chromatography. Polyclonal antibodies were produced in mice against the recombinant protein and the antibodies reacted specifically with the recombinant protein and XSV in WTD-infected tissues. This is the first report of detection of XSV using antibodies against recombinant capsid protein.

Genomic and antibody-based assays for the detection of Indian strains of Macrobrachium rosenbergii nodavirus and extra small virus associated with white tail disease of Macrobrachium rosenbergii

“…Available antibody-based diagnostic methods for MrNV includes S-ELISA [30] and TAS-ELISA [26] and for XSV, it is polyclonal antibodies raised against the whole virus [33] and recombinant viral protein [21]. In the majority of cases, antibodies were produced against the whole virus.…”

Recombinant viral proteins delivered orally through inactivated bacterial cells induce protection in Macrobrachium rosenbergii (de Man) against White Tail Disease

“…The genome of Mr NV consists of two single‐stranded positive‐sense RNA molecules encoding RNA‐dependent RNA polymerase (RdRp), B2 protein and a capsid protein (Bonami et al., 2005; Hayakijkosol et al., 2012; Low et al., 2018; NaveenKumar, 2017; NaveenKumar, Karunasagar et al., 2013; NaveenKumar, Shekar, et al., 2013). The smaller virus among the two XSV is also non‐enveloped, 14–16 nm diameter with a single‐stranded positive‐sense RNA genome of approximately 0.8 kilobases (Qian et al., 2003; Sri Widada & Bonami, 2004) which codes for two overlapping structural (capsid) proteins of size 16 and 17 kDa (Neethi et al., 2012; Wang et al., 2008). Due to its small size and absence of RdRp or enzymes required for replication, it has been suggested that XSV may be a satellite virus and replication depends on its helper virus, Mr NV in this case (Sri Widada & Bonami, 2004; Zhang et al., 2006).…”