Peptide-recombinant VP6 protein based enzyme immunoassay for the detection of group A rotaviruses in multiple host species

Naveen Kumar, Yashpal Singh Malik, Satish Kumar, Kuldeep Sharma, Subhankar Sircar, Sharad Saurabh, Baldev R. Gulati, Neeraj Singh, Arvind Kumar Singh, Vinay G. Joshi, K. Bányai, Kuldeep Dhama

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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.

Original languageEnglish
Article numbere0159027
JournalPLoS One
Volume11
Issue number7
DOIs
Publication statusPublished - Jul 1 2016

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ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Kumar, N., Malik, Y. S., Kumar, S., Sharma, K., Sircar, S., Saurabh, S., Gulati, B. R., Singh, N., Singh, A. K., Joshi, V. G., Bányai, K., & Dhama, K. (2016). Peptide-recombinant VP6 protein based enzyme immunoassay for the detection of group A rotaviruses in multiple host species. PLoS One, 11(7), [e0159027]. https://doi.org/10.1371/journal.pone.0159027