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.

Highlights

  • Rotaviruses (RVs) belong to the family Reoviridae and are a leading cause of diarrhea in humans and animals worldwide

  • We identified six regions of high antigenic index within bovine RVA VP6 protein and correspondingly synthesized peptides by solid phase peptide synthesis over Wang resin

  • The Circular Dichroism (CD) spectra analysis revealed that all six peptides in polar solvent had a negative minimum below 200 nm in a far UV range

Read more

Summary

Introduction

Rotaviruses (RVs) belong to the family Reoviridae and are a leading cause of diarrhea in humans and animals worldwide. RVs account for approximately one fourth of global mortality in Indian children annually [1,2,3]. RVs are classified into at least nine distinct serological species or groups (A to I), of which group A rotaviruses (RVAs) are frequently associated with acute diarrhea [1,2,5]. Used dual classification system for RVAs designate P- and G-genotypes to the genes coding for outer most proteins VP4 and VP7, respectively. At least 27 G and 37 P genotypes of RVAs have been identified in humans and animals, reflecting huge genetic diversity among RVs [2,6]

Objectives
Methods
Results
Conclusion
Full Text
Published version (Free)

Talk to us

Join us for a 30 min session where you can share your feedback and ask us any queries you have

Schedule a call