Competitive ELISA for the Detection of Antibodies to Rift Valley Fever Virus in Goats and Cattle

Abstract

ABSTRACT. Rift Valley fever virus (RVFV) is one of the important emerging viral diseases of serious impact in public health and animal hygiene both in human and animal industries. In this study, we developed a monoclonal antibody-based competitive ELISA for the detection of antibodies to RVFV in goats and cattle. The recombinant N protein of RVFV was expressed in E. coli with a six-histidine tag, and the purified N protein was used for detecting antigen with a competitive monoclonal antibody against RVFV antibodies. The competitive ELISA (C-ELISA) could detect antibodies at 9-11 days after inoculation in goats and cattle with a sensitivity of 94.7% (virus neutralization titer >32) and specificity of 99.7%, respectively. In addition, the C-ELISA did not show any cross-reactivity with positive sera against arboviruses such as Akabane, Aino, Chuzan, Ibaraki and bovine ephemeral fever virus, which are prevalent viral agents in ruminant animals throughout Southeast Asia. The results of the present study indicate that the C-ELISA is a simple, rapid and convenient serodiagnostic method for RVFV in goats and cattle.

Large-Scale International Validation of an Indirect ELISA Based on Recombinant Nucleocapsid Protein of Rift Valley Fever Virus for the Detection of IgG Antibody in Domestic Ruminants

“…The diagnostic accuracy estimates determined in our study are similar to those previously published for ELISAs based on a whole RVFV antigen [36][37][38][39][40][41]55] as well as those based on recombinant NP ELISAs [51][52][53][54][55][56][57][58][59][60], including the commercially available NP-based competition ELISAs [52,60]. Although recorded at a relatively low rate, false-positive results were documented in this study across different subpopulations and are of particular concern in ruminants originating from RVF-non-endemic countries.…”

“…The inhibition ELISA based on a whole tissue culture-derived, inactivated antigen [ 40 ] and competitive ELISA based on recombinant NP antigen [ 50 , 51 , 52 ] allow multi-species RVFV antibody detection using the same diagnostic procedure without requirements for species-specific conjugates. While in the last 15 years, several groups have developed and evaluated various ELISA formats based on RVFV recombinant NP antigen [ 43 , 44 , 45 , 46 , 47 , 48 , 49 , 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 , 59 , 60 ], more extensive evaluation of their diagnostic performance was only achieved in humans [ 58 ], buffalo [ 59 ], and cattle [ 60 ] to date.…”

Validation of a Commercially Available Indirect Elisa Using a Nucleocapside Recombinant Protein for Detection of Schmallenberg Virus Antibodies

“…Some cross reactions between nucleocapsid protein and IgG antibodies induced by infection with other Simbu serogroup virus members might occur [14]. Limited cross reactions were observed with sera from ruminants experimentally infected with Akabane virus (AKAV) (data not shown).…”

“…Using the same strategy developed for Rift valley fever ELISA, the N viral protein of SBV has been expressed in a prokaryote system with a 6 histidine tag [14]. After expression, denaturation in the presence of urea, purification by IMAC chromatography and some desalting steps, the SBV N recombinant protein was used as antigen to discriminate sera from SBV infected from naive animals.…”

Rift Valley Fever Virus, Japanese Encephalitis Virus, and African Swine Fever Virus: Three Transboundary, Vector-Borne, Veterinary Biothreats With Diverse Surveillance, and Response Capacity Needs

“…VanVuren and Paweska developed an ELISA based on the capture of the nucleocapsid protein (NP) (57). Alternatively, a monoclonal antibody-based competitive ELISA developed by Kim et al for the detected antibodies against RVFV in goats and cattle with a sensitivity/specificity of 94.7/99.7% between 9 and 11 days post-inoculation (58). A multiplex fluorescence microsphere immunoassay (FMIA) also detected cattle and sheep IgM and IgG antibodies to the RVFV glycoprotein, nonstructural proteins, and nucleoprotein (59).…”