title: Peptide-Recombinant VP6 Protein Based Enzyme Immunoassay for the Detection of Group A Rotaviruses in Multiple Host Species.
abstract: We developed a novel enzyme immunoassay for the detection of group A rotavirus (RVA) antigen in fecal samples of multiple host species. The assay is based on the detection of conserved VP6 protein using anti-recombinant VP6 antibodies as capture antibodies and anti-multiple antigenic peptide (identified and constructed from highly immunodominant epitopes within VP6 protein) antibodies as detector antibodies. The clinical utility of the assay was evaluated using a panel of 914 diarrhoeic fecal samples from four different host species (bovine, porcine, poultry and human) collected from diverse geographical locations in India. Using VP6- based reverse transcription-polymerase chain reaction (RT-PCR) as the gold standard, we found that the diagnostic sensitivity (DSn) and specificity (DSp) of the new assay was high [bovine (DSn = 94.2% & DSp = 100%); porcine (DSn = 94.6% & DSp = 93.3%); poultry (DSn = 74.2% & DSp = 97.7%) and human (DSn = 82.1% & DSp = 98.7%)]. The concordance with RT-PCR was also high [weighted kappa (k) = 0.831-0.956 at 95% CI = 0.711-1.0] as compared to RNA-polyacrylamide gel electrophoresis (RNA-PAGE). The performance characteristics of the new immunoassay were comparable to those of the two commercially available ELISA kits. Our results suggest that this peptide-recombinant protein based assay may serve as a preliminary assay for epidemiological surveillance of RVA antigen and for evaluation of vaccine effectiveness especially in low and middle income settings.
authors.affiliation: Biological Standardization Division, Indian Veterinary Research Institute (IVRI), Izatnagar, Uttar Pradesh, India.
authors.forename: Naveen
authors.initials: N
authors.lastname: Kumar
authors.affiliation: Biological Standardization Division, Indian Veterinary Research Institute (IVRI), Izatnagar, Uttar Pradesh, India.
authors.forename: Yashpal Singh
authors.initials: YS
authors.lastname: Malik
authors.affiliation: Division of Animal Biotechnology, Indian Veterinary Research Institute (IVRI), Izatnagar, Uttar Pradesh, India.
authors.forename: Satish
authors.initials: S
authors.lastname: Kumar
authors.affiliation: National Institute of Research in Tribal Health, Jabalpur, Madhya Pradesh, India.
authors.forename: Kuldeep
authors.initials: K
authors.lastname: Sharma
authors.affiliation: Biological Standardization Division, Indian Veterinary Research Institute (IVRI), Izatnagar, Uttar Pradesh, India.
authors.forename: Subhankar
authors.initials: S
authors.lastname: Sircar
authors.affiliation: Biological Standardization Division, Indian Veterinary Research Institute (IVRI), Izatnagar, Uttar Pradesh, India.
authors.forename: Sharad
authors.initials: S
authors.lastname: Saurabh
authors.affiliation: National Research Center on Equines (NRCE), Hisar, Haryana, India.
authors.forename: Baldev R
authors.initials: BR
authors.lastname: Gulati
authors.affiliation: Biological Standardization Division, Indian Veterinary Research Institute (IVRI), Izatnagar, Uttar Pradesh, India.
authors.forename: Neeraj
authors.initials: N
authors.lastname: Singh
authors.affiliation: Division of Animal Biotechnology, Indian Veterinary Research Institute (IVRI), Izatnagar, Uttar Pradesh, India.
authors.forename: Arvind Kumar
authors.initials: AK
authors.lastname: Singh
authors.affiliation: Division of Animal Biotechnology, Indian Veterinary Research Institute (IVRI), Izatnagar, Uttar Pradesh, India.
authors.forename: Vinay G
authors.initials: VG
authors.lastname: Joshi
authors.affiliation: Institute for Veterinary Medical Research, Centre for Agricultural Research, Hungarian Academy of Sciences, Hungáriakrt. 21, Budapest, Hungary.
authors.forename: Krisztian
authors.initials: K
authors.lastname: Banyai
authors.affiliation: Division of Veterinary Immunology, Indian Veterinary Research Institute (IVRI), Izatnagar, Uttar Pradesh, India.
authors.forename: Kuldeep
authors.initials: K
authors.lastname: Dhama
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology*; D000956:Antigens, Viral / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D002417:Cattle; D002418:Cattle Diseases / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006168:Guinea Pigs; D006801:Humans; D011200:Poultry; D011201:Poultry Diseases / Q000276:immunology*; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D012401:Rotavirus / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D012400:Rotavirus Infections / Q000276:immunology / Q000662:veterinary
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D011994:Recombinant Proteins; C061332:VP6 protein, Rotavirus