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Research (Published online: 07-04-2015)

2. Diagnosis of peste des petits ruminants infection in small ruminants through in-house developed Indirect ELISA: Practical considerations - K. K. Sharma, D. P. Kshirsagar, I. H. Kalyani, D. R. Patel, P. D. Vihol and J. M. Patel

Veterinary World, 8(4): 443-448

 

 

   doi: 10.14202/vetworld.2015.443-448

 

K. K. Sharma: Department of Veterinary Microbiology, Vanbandhu College of Veterinary Science and Animal Husbandry, Navsari Agricultural University, Navsari, Gujarat, India; kishan12sharma@rediffmail.com

D. P. Kshirsagar: Department of Veterinary Public Health and Epidemiology, Vanbandhu College of Veterinary Science and Animal Husbandry, Navsari Agricultural University, Navsari, Gujarat, India; drdpk04v@gmail.com

I. H. Kalyani: Department of Veterinary Microbiology, Vanbandhu College of Veterinary Science and Animal Husbandry, Navsari Agricultural University, Navsari, Gujarat, India; ihkvet@gmail.com

D. R. Patel: Department of Veterinary Microbiology, Vanbandhu College of Veterinary Science and Animal Husbandry, Navsari Agricultural University, Navsari, Gujarat, India; vetdharmesh74@yahoo.com

P. D. Vihol: Department of Veterinary Pathology, Vanbandhu College of Veterinary Science and Animal Husbandry, Navsari Agricultural University, Navsari, Gujarat, India; drpriti@nau.in

J. M. Patel: Department of Veterinary Pathology, Vanbandhu College of Veterinary Science and Animal Husbandry, Navsari Agricultural University, Navsari, Gujarat, India; dr.jams@rediffmail.com

 

Received: 26-11-2014, Revised: 25-02-2015, Accepted: 05-03-2015, Published online: 07-04-2015

 

Corresponding author: D. P. Kshirsagar, e-mail: drdpk04v@gmail.com


Citation: Sharma KK, Kshirsagar DP, Kalyani IH, Patel DR, Vihol PD, Patel JM (2015) Diagnosis of peste des petits ruminants infection in small ruminants through in-house developed Indirect ELISA: Practical considerations, Veterinary World 8(4): 443-448.



Aim: The work was conducted to diagnose peste des petits ruminants (PPR) outbreak through an in house developed indirect ELISA (thereafter referred as iELISA) its comparison with other available diagnostic tests and description of practical considerations in its development, utility and limitations.

Materials and Methods: An outbreak resembled to PPR occurred in two different places of southern Gujarat viz. Vapi and Navsari, affecting 622 animals, including both goat (n = 476) and sheep (n = 146). Animals displayed the typical signs of PPR at Vapi; however diarrhea was the inconsistent feature in animals of Navsari. The affection caused morbidity of 100% and mortality were 73.68% (n = 392/532) and 56.67% (n = 51/90) in Vapi and Navsari outbreaks, respectively. Relevant ante mortem and post mortem samples were collected from representative animals. At the outset of the epidemic no kit was available with us, so agar gel immunodiffusion (AGID) was carried out and a commercial ELISA (cELISA) kit was ordered for making diagnosis through antibody demonstration. Meanwhile, an iELISA was developed in house using PPR vaccine as antigen and protein G conjugated HRPO antibody as detector. Histopathology and results of sandwich ELISA were also used to diagnose PPR virus (PPRV) in the outbreak.

Results: The iELISA developed had detected PPRV antibodies in 22/24 samples (91.66%). Significant difference was observed in disease sensitivity pattern of two species by Chi-square test. While AGID failed to detect antibodies in any sample. Results were reconfirmed by comparing with commercially available cELISA kit.

Conclusion: PPR is an economically important disease and for the rapid diagnosis of PPR the in house developed antibody capture iELISA can be a suitable cost effective alternative.

Keywords: agar gel immunodiffusion, competitive ELISA, Goat, indirect ELISA, peste des petits ruminants.



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