Avidin-Biotin recombinant nucleoprotein competitive ELISA for the detection of peste des petits ruminants virus antibodies in sheep and goats.

Journal of Virological Methods
V BalamuruganB R Shome

Abstract

The present study describes the development of a truncated recombinant peste des petits ruminants virus (PPRV) nucleoprotein (rPPRV-NPN) and its polyclonal antibodies-based immuno-diagnostic assay, Avidin-Biotin (AB) recombinant nucleoprotein competitive ELISA (ABrC-ELISA) for the detection of PPRV antibodies in the sheep and goats. The PPRV N-terminal immunogenic region (1-266 aa) of nucleoprotein (NPN) coding sequence was amplified and cloned into the pETite vector. The rPPRV-NPN with a molecular weight of ∼ 30 kDa was expressed in E. coli, purified, and characterized by SDS-PAGE and immunoblot using standard PPRV specific sera. The Ni-NTA affinity-purified rPPRV-NPN as coating antigen and its hyperimmune serum as competitive antibodies raised in guinea pigs were evaluated as diagnostic reagents in ABrC-ELISA using the known standard panel of sera. The threshold (cut-off) Percentage Inhibition (PI) value was determined as 45 (mean ± 3 SD) based on the reactivity of the known sheep and goats sera to PPRV antibodies [negative (n = 140) and positive (n = 98)] and the assay had a sensitivity of 97 % (95 % Confidence Interval (CI): 91.3-99.4 %) and specificity of 100 % (95 % CI: 97.4-100 %) with an excellent Area under curve (AUC)...Continue Reading

References

Feb 2, 1991·The Veterinary Record·P C LefèvreG Staak
Aug 26, 1998·Revue Scientifique Et Technique·R H Jacobson
Aug 26, 1998·Revue Scientifique Et Technique·R S Schrijver, J A Kramps
Dec 2, 2006·Veterinary Research Communications·D MuthuchelvanS K Bandyopadhyay
Oct 6, 2015·Virusdisease·Gnanavel VenkatesanBimelendu Mondal
Jul 22, 2018·Biologicals : Journal of the International Association of Biological Standardization·Suresh H BasagoudanavarRamamurthy Venkataramanan

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