Please cite this article in press as: Yousuf, R.W., et al., Development of a single-plate combined indirect ELISA (CI-ELISA) for the detection of antibodies against peste-des-petits-ruminants and bluetongue viruses in goats. Small Ruminant Res. (2015), http://dx.doi.org/10.1016/j.smallrumres.2015.01.007 ARTICLE IN PRESS G Model RUMIN-4860; No. of Pages 3 Small Ruminant Research xxx (2015) xxx–xxx Contents lists available at ScienceDirect Small Ruminant Research jou r n al homep age : w w w . elsevier.com/locate/smallrumres Short communication Development of a single-plate combined indirect ELISA (CI-ELISA) for the detection of antibodies against peste-des-petits-ruminants and bluetongue viruses in goats Raja Wasim Yousuf a , Arnab Sen b , Bimalendu Mondal a , Sanchay Kumar Biswas a , Karam Chand a , Kaushal Kishore Rajak a , Gopal R. Gowane c , Shashi Bhusan Sudhakar a , Awadh Bihari Pandey a , Muthannan Andavar Ramakrishnan a , Dhanavelu Muthuchelvan a,∗ a Division of Virology, Indian Veterinary Research Institute, Mukteswar Campus, Uttarakhand 263 138, India b Division of Animal Health, ICAR Research Complex for N.E.H Region, Umiam, Meghalaya 193 103, India c Central Sheep and Wool Research Institute, Avikanagar, Rajasthan 304 501, India a r t i c l e i n f o Article history: Received 2 November 2013 Received in revised form 11 January 2015 Accepted 14 January 2015 Available online xxx Keywords: Indirect-ELISA c-ELISA Single plate combined ELISA Peste-des-petits-ruminants Bluetongue Goat a b s t r a c t Peste-des-petits-ruminants and bluetongue are two of the most important viral diseases of small ruminants. In the present study, a combined indirect enzyme-linked immunosorbent assay (CI-ELISA) was developed, which can be run parallelly on the same plate for simul- taneous detection of antibodies against peste-des-petits-ruminants virus and bluetongue virus. A total of 503 goat sera samples were used for developing the CI-ELISA. The relative sensitivity and specificity of CI-ELISA for detection of antibodies against PPRV were 90% and 88.8%, respectively at cut off level of 34.52% positivity. In case of BTV, 95.9% relative sensi- tivity and 98.6% specificity values were obtained at cut-off level of 40.42% positivity. These results indicate that the single plate CI-ELISA reported here could be used as an alternate method to existing dual plate competitive ELISAs for diagnosis/sero-surveillance of both the diseases. © 2015 Elsevier B.V. All rights reserved. 1. Introduction Peste-des-petits-ruminants (PPR) and bluetongue (BT) are two of the most important viral diseases of small rumi- nants with serious socio-economic impact. PPR is caused by peste-des-petits-ruminants virus (PPRV), which belongs to the genus Morbillivirus of family Paramyxoviridae, and BT is caused by bluetongue virus (BTV) of the genus Orbivirus of family Reoviridae (Van Regenmortel et al., 2000). Both the diseases are endemic in India and a national control ∗ Corresponding author. Tel.: +91 5942286346; fax: +915942286347. E-mail address: drchelva@gmail.com (D. Muthuchelvan). program (NCP-PPR) has been recently launched by Indian government (Annual Report, 2012–13) for control of PPR. However, BT vaccine is yet to be available in the market. PPRV infection in goats can lead to marked suppression of host immune response accompanied by severe leuco- penia which may increase the incidence and severity of other diseases (Rajak et al., 2005). BT is vector-borne, noti- fiable disease and occurs as subclinical form of the disease in goats (Maclachlan et al., 2009). Mixed infection of PPRV with BTV (Mondal et al., 2009), orf virus (Saravanan et al., 2007) and goatpox virus (Malik et al., 2011) have been reported. In India, diagnosis/sero-surveillance of PPR has been carried out using monoclonal antibody based c-ELISA (Singh et al., 2004) or polyclonal antibody based indirect http://dx.doi.org/10.1016/j.smallrumres.2015.01.007 0921-4488/© 2015 Elsevier B.V. All rights reserved.