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			<p class="Type-of-Article">Review Article</p>
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			<p class="title- ParaOverride-1">&nbsp;</p>
  <p class="title- ParaOverride-1">Peste-Des-Petits-Ruminants: An Indian Perspective</p>
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			<p class="Authors ParaOverride-1">&nbsp;</p>
			<p class="Authors ParaOverride-1"><span class="CharOverride-1">Dhanavelu Muthuchelvan</span><span class="CharOverride-2">1*</span><span class="CharOverride-1">, Kaushal Kishor Rajak</span><span class="CharOverride-2">1</span><span class="CharOverride-1">, Muthannan Andavar Ramakrishnan</span><span class="CharOverride-2">1</span><span class="CharOverride-1">, Dheeraj Choudhary</span><span class="CharOverride-2">1</span><span class="CharOverride-1">, Sakshi Bhadouriya</span><span class="CharOverride-2">1</span><span class="CharOverride-1">,</span><span class="CharOverride-2"> </span><span class="CharOverride-1">Paramasivam Saravanan</span><span class="CharOverride-2">2</span><span class="CharOverride-1">, Awadh Bihari Pandey</span><span class="CharOverride-2">1</span><span class="CharOverride-1">, Raj Kumar Singh</span><span class="CharOverride-2">3</span></p>
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			<p class="Affiliations ParaOverride-1"><span class="CharOverride-3">1</span>Division of Virology, Indian Veterinary Research Institute, Mukteswar Campus, Nainital, Uttarakhand 263 138, India; <span class="CharOverride-3">2</span>Indian Veterinary Research Institute, Hebbal Bengaluru, 560024, Karnataka, India; <span class="CharOverride-3">3</span>Indian Veterinary Research Institute, Izatnagar, 243122, India.</p>
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			<p class="Abstract ParaOverride-1">&nbsp;</p>
			<p class="Abstract ParaOverride-1"><span class="Abstract CharOverride-4">Abstract</span><span class="Abstract"> | </span><span class="Abstract" xml:lang="en-US">Peste-des-petits-ruminants (PPR) is an acute or subacute, highly contagious viral disease of small ruminants, characterized by fever, oculonasal discharges, stomatitis, diarrhoea and pneumonia with high morbidity and mortality. Peste-des-petits-ruminants virus (PPRV), the etiological agent of PPR, is antigenically related to another rinderpest virus (RP) which was globally eradicated. PPR is gaining worldwide attention through the concerted effort of scientists working together under the aegis of global PPR research alliance (GPRA). The first homologous live attenuated vaccine was developed using Nigeria 75/1, which has been used worldwide. In India, live attenuated vaccines have been developed using Sungri 96, Arasur 87 and Coimbatore 97 viruses. In this review, the status of PPR and control strategy with special reference to the Indian context is comprehensively discussed.</span></p>
		  <p class="Abstract ParaOverride-1">&nbsp;</p>
			<p class="Abstract ParaOverride-1"><span class="Abstract CharOverride-4">Keywords</span><span class="Abstract"> | </span><span class="Abstract" xml:lang="en-US">PPR, PPRV, Vaccine, DIVA, Eradication, Symptoms, Epidemiology, Diagnosis, Vaccines, Immunity, Control programme, Replication</span></p>
		  <p class="Abstract ParaOverride-1">&nbsp;</p>
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			<p class="Editor----Citation"><span class="CharOverride-4">Editor</span> | Muhammad Munir (DVM, PhD), Avian Viral Diseases Program, Compton Laboratory, Newbury, Berkshire, RG20 7NN, UK.</p>
			<p class="Editor----Citation"><span class="CharOverride-4">Received</span> | April 27, 2015; <span class="CharOverride-4">Revised</span> | June 16, 2015; <span class="CharOverride-4">Accepted</span> | June 18, 2015; <span class="CharOverride-4">Published</span> | June 24, 2015&#9;&#9;</p>
			<p class="Editor----Citation"><span class="CharOverride-4">*Correspondence</span> | Dhanavelu Muthuchelvan, Indian Veterinary Research Institute, Nainital, Uttarakhand, India; <span class="CharOverride-4">Email:</span> drchelva@gmail.com</p>
			<p class="Editor----Citation"><span class="CharOverride-4">Citation</span> | Muthuchelvan D, Rajak KK, Ramakrishnan MA, Choudhary D, Bhadouriya S, Saravanan P, Pandey AB, Singh RK (2015). Peste-des-petits-ruminants: An Indian perspective. Adv. Anim. Vet. Sci. 3(8): 422-429.  </p>
			<p class="Editor----Citation"><span class="CharOverride-4">DOI</span> | <a href="http://dx.doi.org/10.14737/journal.aavs/2015/3.8.422.429"><span class="Hyperlink">http://dx.doi.org/10.14737/journal.aavs/2015/3.8.422.429</span></a></p>
			<p class="Editor----Citation"><span class="Editor---Citation CharOverride-4">ISSN (Online)</span> | 2307-8316; <span class="Editor---Citation CharOverride-4">ISSN (Print)</span> | 2309-3331</p>
			<p class="Editor----Citation"><span class="CharOverride-4">Copyright</span> © 2015 Muthuchelvan et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</p>
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			<p class="Heading-1--Introduction----">&nbsp;</p>
		  <p class="Heading-1--Introduction----">Introduction</p>
			<p class="Caps-on-First-Para ParaOverride-1">&nbsp;</p>
		  <p class="Caps-on-First-Para ParaOverride-1"><span class="_idGenDropcap-1">P</span><span>este-des-petits-ruminants </span>(PPR) is a viral disease of sheep and goats caused by PPR virus (PPRV) with huge economical concern. India harbours 65.06 and 135.17 million of sheep and goats, respectively (19<span class="CharOverride-7">th</span> Livestock census, 2012; <a href="http://dahd.nic.in/dahd/WriteReadData/Livestock.pdf"><span class="Hyperlink">http://dahd.nic.in/dahd/WriteReadData/Livestock.pdf</span></a>) which is 16.1 and 6.4 % of the world’s total goat and sheep population, respectively. The disease threatens 70% of the landless labourers, small and medium farmers who rears sheep and goats for their livelihood. As the disease affects the poor sections of the society, international agencies such as FAO &amp; OIE set a target for global eradication by 2030 (<a href="http://www.fao.org/ppr/en/"><span class="Hyperlink">http://www.fao.org/ppr/en/</span></a>). The disease causes up to 100% mortality in and productive loss. It is estimated that PPR alone leads to annual losses of 1,800 million Indian Rupees (US$ 39 million) (<a href="#Singh-RP--2012"><span class="Hyperlink">Singh, 2012</span></a>). As one of the forerunners in the rinderpest (RP) research and significant contribution in the global eradication of the disease, consequently, our lab developed live attenuated vaccine (Sungri96) and monoclonal antibody based diagnostics (<a href="#Singh-RP--Sreeniva-2004b"><span class="Hyperlink">Singh et al., 2004b</span></a>, <a href="#Singh-RP--Sreenivasa-2004c"><span class="Hyperlink">c</span></a>). The vaccine and diagnostics are supplied throughout India as well as neighbouring countries. Availability of these technologies helped India to initiate mass vaccination campaign which brought down the incidence of the disease significantly (<a href="#Figure-1-"><span class="Hyperlink">Figure 1</span></a>). Further, Government of India has recently launched a Peste des Petits Ruminants Control Programme (PPR-CP). The PPR Control Programme involving intensive vaccination of susceptible animals has been started in 2010. The programme involves vaccinating all susceptible goats and sheep and three subsequent generations. Under first phase, States of Kerala, Tamil Nadu, Karnataka, Andhra Pradesh, Maharashtra, Goa and UTs of Lakshadweep, Daman and Diu, Dadra and Nagar Haveli, Andaman and Nicobar Islands and Pondicherry were covered. In the second phase, the programme has been expanded to all States/UTs in February, 2014 <a href="http://dahd.nic.in/dahd/WriteReadData/Animal%20Husbandry%20English%202014-15%20(1)"><span class="Hyperlink">http://dahd.nic.in/dahd/WriteReadData/Animal%20Husbandry%20English%202014-15%20(1)</span></a>.<span class="Hyperlink">pdf</span>. This short review focuses on the current scenario of PPR in India. </p>
			<p class="Heading-1--Introduction----">&nbsp;</p>
		  <p class="Heading-1--Introduction----">The Virus</p>
			<p class="Body-Text">&nbsp;</p>
		  <p class="Body-Text">Peste-des-petits-ruminants virus (PPRV) is a member of the genus <span class="CharOverride-8">Morbillivirus. </span>subfamily <span class="CharOverride-8">Paramyxovirinae, </span>family <span class="CharOverride-8">Paramyxoviridae </span>in the order <span class="CharOverride-8">Mononegavirales </span>(<a href="#International-Committee-on-Taxonom"><span class="Hyperlink">International Committee on Taxonomy of Viruses, 2012</span></a>). Since the virus is enveloped one, it is easily inactivated under sunlight and with many chemicals. The genome of PPRV is a negative sense, single-stranded RNA with the size of 15948 bp. The genome is organized into six transcriptional units and each encodes at least one non overlapping protein: the nucleocapsid (N), the matrix (M), the polymerase or large (L), the phosphoprotein (P), and two envelope proteins haemagglutin (H) and fusion (F). The P protein uses alternate expression strategies to code for two non-structural proteins <span class="CharOverride-8">viz</span>., V and C (<a href="#Muthuchelvan-D--Sa-2006"><span class="Hyperlink">Muthuchelvan et al., 2006</span></a>). The PPRV genes arranged from 3<span class="CharOverride-9">ˈ</span> to 5’ on the genome is in an order of N-P-M-F-H-L separated by intergenic region which is CTT in most cases. At the 3’ and 5<span class="CharOverride-9">ˈ </span>end of the genome there is a leader (52 nucleotides) and trailer (37 nucleotides) region contain promoter functions (<a href="#Bailey-D--Banyard-A--Dash-P--Ozkul-A--Barrett-T--2005"><span class="Hyperlink">Bailey et al., 2005</span></a>). Two heptad repeats (HR1 and HR2) of the fusion protein was shown to form a six-helix and trimeric coiled-coil bundle for initiation of fusion activity (<a href="#Rahaman-A--Srinivasan-N--Shamala-N--Sh"><span class="Hyperlink">Rahaman et al., 2003</span></a>). Host-pathogen interaction study with vaccine virus (Sungri96) and goat peripheral blood mononuclear cells (PBMCs) revealed the dysregulation of immune regulatory pathways (<a href="#Manjunath-S--Kumar-G--Mishra-B--Mishra-B-"><span class="Hyperlink">Manjunath et al., 2015</span></a>). Establishment of an <span class="CharOverride-8">in vitro</span> system using Sungri96 virus shown that RNP complex is actively synthesis mRNA (<a href="#Yunus-M--Shaila-MS--2012-."><span class="Hyperlink">Yunus and Shaila, 2012</span></a>). PPRV multiplication was reduced significantly in <span class="CharOverride-10">S</span>ignaling <span class="CharOverride-10">L</span>ymphocyte <span class="CharOverride-10">A</span>ctivation <span class="CharOverride-10">M</span>olecule (SLAM;CD150) receptors suppressed with siRNA in B95a (<a href="#Pawar-RM--Raj-GD--2008b"><span class="Hyperlink">Pawar et al., 2008b</span></a>).Transiently expressed PPRV F glycoprotein induces cell fusion in the absence of H protein (<a href="#Seth-S--Shaila-MS--2001"><span class="Hyperlink">Seth and Shaila, 2001</span></a>). The H protein was found to have potential T cell determinant(s) at amino acids position 123-137 and 242-609 (<a href="#Sinnathamby-G--Renukaradhya-GJ--Rajasekhar-M--Nayak-R--Shaila-MS--2001"><span class="Hyperlink">Sinnathamby et al., 2001</span></a>). </p>
			<p class="Heading-1--Introduction----">&nbsp;</p>
		  <p class="Heading-1--Introduction----">The disease</p>
			<p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">PPR is a disease of small ruminants which resembles rinderpest of cattle. The incubation period is 3-6 days, followed by high fever, oculonasal discharges, pneumonia, stomatitis, and inflammation of gastrointestinal tract leading to severe diarrhoea followed by death or recovery (<a href="#Balamurugan-V-2014a"><span class="Hyperlink">Balamurugan et al., 2014a</span></a>; <a href="#Sen-A--Saravanan-2010"><span class="Hyperlink">Sen et al., 2010</span></a>; <a href="#Zahur-AB--Irshad-H--Hussain-M--Ullah-A--Jahangir-M--et-al.--2008"><span class="Hyperlink">Zahur et al., 2008</span></a>). The disease causes more severe lesions in goats than sheep. Although, the reason for this host specificity is not fully understood, difference in genetic makeup and/or receptor distributions of the host might have a role. Different levels of SLAM mRNA could influence the virus replication in different species (<a href="#Pawar-RM--Dhinaka-2008a"><span class="Hyperlink">Pawar et al., 2008a</span></a>). Another study examined the replication of PPRV in PBMCs of Indian goats and water buffalo and demonstrated that the level of TLR3 and TLR7 and downstream signalling molecules correlate with susceptibility (<a href="#Dhanasekaran-S--Biswas-M--Vignesh-A"><span class="Hyperlink">Dhanasekaran et al., 2014</span></a>). Incidence in other species such as cattle, pig, camel, buffalo, lion and captive wild small ruminants were reported but are not contributing to the disease epidemiology. Experimental infection of cattle with PPRV revealed that the nucleic acid could be detected up to 397 days post infection (<a href="#Sen-A--Saravanan-P"><span class="Hyperlink">Sen et al., 2014</span></a>). </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">No carrier state or persistent infections have been reported, however, disease may circulate in subclinical forms in sheep and goats of endemic region. The recovered animals are immune for lifelong. The cytokine expression profile (IL-4 and IFN-<span class="CharOverride-9">γ</span>) of vaccinated and infected goats were found to have a unique biphasic response of IL-4 expression with up-regulation of IFN-<span class="CharOverride-9">γ</span> on 7<span class="CharOverride-7">th</span> days post vaccination (<a href="#Patel-A--Rajak-KK--Balamurugan-V--Sen-A--Sudhakar-SB--et-al.--2012-.-Cytokines-expres"><span class="Hyperlink">Patel et al., 2012</span></a>). PPRV was reported to induce apoptosis <span class="CharOverride-8">in vitro</span> in goat PBMCs (<a href="#Mondal--Sreenivasa--Dh-2001"><span class="Hyperlink">Mondal et al., 2001</span></a>).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			
            <div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150625033042.png" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150625033042.png" width="80" height="80"></a>
            
          <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-4"><a id="Figure-1-"></a>Figure 1: </span><a href="http://nexusacademicpublishers.com/uploads/figures/20150625033042.png">The disease outbreak status for the period between 2009 and 2014</a></p>
       </div>

			
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
		  <p class="Body-Text ParaOverride-1">Although, presence of PPR-like disease has been suspected earlier in retrospective study (<a href="#Taylor-WP--Diallo-A--Gopalakrishna-S--Sreeramalu-P--Wilsmore-AJ--et-al.--2002"><span class="Hyperlink">Taylor et al., 2002</span></a>), its presence was confirmed in India in1987 from Arasur village of Villupuram district of Tamil Nadu (<a href="#Shaila-MS--Purush-1989"><span class="Hyperlink">Shaila et al., 1989</span></a>). Currently, PPR outbreaks are being reported regularly from different parts of the country (<a href="#Chauhan-HC--Lambade-PS--Sen-A--Dadawala-AI--Ranaware-PB--et-al.--2011"><span class="Hyperlink">Chauhan et al., 2011</span></a>; <a href="#Kerur-N--Jhala-MK--Joshi-CG--200"><span class="Hyperlink">Kerur et al., 2008</span></a>; <a href="#Muthuchelvan-D--De-A-2014"><span class="Hyperlink">Muthuchelvan et al., 2014</span></a>; <a href="#Nanda-YP--Chatterjee-A--Purohit-AK--Diallo-A--Innui-K--et-al.--1996-.-The-isolati"><span class="Hyperlink">Nanda et al., 1996</span></a>; <a href="#Raghavendra-AG--Gajendragad-M"><span class="Hyperlink">Raghavendra et al., 2008</span></a>; <a href="#Singh-RP--Saravanan-2004a"><span class="Hyperlink">Singh et al., 2004a</span></a>). The disease outbreak status for the period between 2009 and 2014 is depicted in <a href="#Figure-1-"><span class="Hyperlink">Figure 1</span></a>. The reported seroprevalence rate of PPRV at country level in goats and sheep was 43.56 % (<a href="#Balamurugan-V--2011"><span class="Hyperlink">Balamurugan et al., 2011</span></a>) and 4.58% in cattle and buffalos (<a href="#Balamurugan-V-2012a"><span class="Hyperlink">Balamurugan et al., 2012a</span></a>). Molecular epidemiological studies confirm the circulation of lineage IV in India (<a href="#Balamurugan-V-2010"><span class="Hyperlink">Balamurugan et al., 2010</span></a>; <a href="#Dhar-P--Sreenivasa-BP--Barrett-T--Cortey"><span class="Hyperlink">Dhar et al., 2002</span></a>; <a href="#Muthuchelvan-D--De-A-2014"><span class="Hyperlink">Muthuchelvan et al., 2014</span></a>; <a href="#Shaila-MS--Shamaki-1996"><span class="Hyperlink">Shaila et al., 1996</span></a>). Presence of mixed infection with bluetongue (<a href="#Mondal--Sen---Cha-2009"><span class="Hyperlink">Mondal et al., 2009</span></a>), goatpox (<a href="#Malik-YS--Singh-D--Chandrashekar-KM"><span class="Hyperlink">Malik et al., 2011</span></a>) and Orf (<a href="#Saravanan-P--Balamurugan-2007"><span class="Hyperlink">Saravanan et al., 2007</span></a>) viruses have also been reported. </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">Disease transmission and Epidemiology</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text">The virus spreads through close contact between infected and susceptible population. The primary portal of entry is via respiratory route. Viraemia develop 2-3 days post infection i.e 1-2 days before the appearance of first clinical sign. Fine infective droplets from the secretions and excretions of the infected animals are released into the air (<a href="#Sen-A--Saravanan-2010"><span class="Hyperlink">Sen et al., 2010</span></a>). Transmission could also occur through contaminated water, feed troughs and bedding (<a href="#Lef-vre-PC--Diallo-A--1990-"><span class="Hyperlink">Lefèvre and Diallo, 1990</span></a>). </p>
		  <p class="Body-Text">&nbsp;</p>
			<p class="Body-Text ParaOverride-1"><span class="CharOverride-12">In India, animals are allowed to share common grazing land and water sources. Besides, migration of animals between various states is common especially, in the sub-Himalayan region and western dry land areas such as Rajasthan and Gujarat </span>(<a href="#Nanda-YP--Chatterjee-A--Purohit-AK--Diallo-A--Innui-K--et-al.--1996-.-The-isolati"><span class="Hyperlink">Nanda et al., 1996</span></a>; <a href="#Singh-RP--Saravanan-2004a"><span class="Hyperlink">Singh et al., 2004a</span></a>)<span class="CharOverride-12">. Mixing of these migrated populations with local population may contribute to the disease transmission. Further, during the festival seasons, animals are shipped to various states for meat purposes. These unrestricted movements of animals contribute significantly to the epidemiology of the disease. In India, the disease occurs round the year and the maximum outbreaks reported during the winter and rainy seasons.&#160;Therefore, vaccination just prior to the onset of rainy/winter season will be more appropriate.</span></p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Initially, PPRV was classified in to 4 lineages I–IV based on the F gene sequencing (<a href="#Dhar-P--Sreenivasa-BP--Barrett-T--Cortey"><span class="Hyperlink">Dhar et al., 2002</span></a>; <a href="#Shaila-MS--Shamaki-1996"><span class="Hyperlink">Shaila et al., 1996</span></a>); lineage I-III viruses were reported in several countries of Africa and lineage IV (Asian lineage) mainly in Middle East and Asia (<a href="#Banyard-AC--Parida-S--Batten-C--Oura-C--Kwiatek-O--Libeau-G--2010"><span class="Hyperlink">Banyard et al., 2010</span></a>; <a href="#Dhar-P--Sreenivasa-BP--Barrett-T--Cortey"><span class="Hyperlink">Dhar et al., 2002</span></a>; <a href="#Ozkul-A--Akca-Y--Alkan-F--Barrett-T--Karaoglu-T"><span class="Hyperlink">Ozkul et al., 2002</span></a>). Currently, N gene is preferred over F gene due to its better molecular separation (<a href="#Kwiatek-O--Minet-C--Grillet-C--Hurard-C--Carlsson-E--et-al.--2007-."><span class="Hyperlink">Kwiatek et al., 2007</span></a>). In the recent past, many researchers reports the presence of lineage IV in African countries (<a href="#Maganga-GD--Verrier-D--Zerbinati-R"><span class="Hyperlink">Maganga et al., 2013</span></a>; <a href="#Salami-H--Croville-G--Kwiatek"><span class="Hyperlink">Salami et al., 2014</span></a>). Till now, only lineage IV viruses have been reported in India. The PPRV goat strain isolated during the recent outbreak at Tripura showed 99.2 to 99.6% nucleotide identities with the Bangladesh strains (<a href="#Muthuchelvan-D--De-A-2014"><span class="Hyperlink">Muthuchelvan et al., 2014</span></a>). The latter study confirms the transboundary transmission of PPRV with the neighbouring countries. </p>
			<p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Heading-1--Introduction----">Vaccine</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Before the development of PPRV vaccine, owing to cross-protection between RPV and PPRV, an attenuated tissue culture RP vaccine (TCRP; Plowright’s strain) was used in many countries. However, since the last phase of RP eradication, this practice was banned. The first homologous vaccine against PPR was developed in 1987 using a Nigerian isolate (Nigeria/75/1) (<a href="#Diallo-A--Mine--Le-2007"><span class="Hyperlink">Diallo et al., 1989, 2007</span></a>). Subsequently, three PPRV vaccines have been developed in India using indigenous isolates of lineage IV (PPRV/Sungri/96, PPRV/Arasur/87, and PPRV/Coimbatore/97) (<a href="#Palaniswami-KS--Thangavelu"><span class="Hyperlink">Palaniswami et al., 2005</span></a>; <a href="#Singh-RP--Saravanan-2004a"><span class="Hyperlink">Singh et al., 2004a</span></a>). Although, all the three vaccines were reported to be equally efficacious (<a href="#Santhosh-AK--Gomes-AR--Hegde-R"><span class="Hyperlink">Santhosh et al., 2013</span></a>; <a href="#Saravanan-P--Sen-2010"><span class="Hyperlink">Saravanan et al. 2010</span></a>), it was observed that the replication cycle and monoclonal reactivity of Arasur/87 virus is different from Sungri/96 virus (<a href="#Singh-RP--De-UK--P-2010"><span class="Hyperlink">Singh et al., 2010</span></a>). Currently, Sungri/96 strain is being used throughout the country. This vaccine is potent, confers life-long immunity and is safer to pregnant animals (<a href="#Rajak-KK--Sreenivasa-BP--Hosamani-M--Singh-RP--Singh-SK--et-al.--2005-.-Experi"><span class="Hyperlink">Rajak et al.</span><span class="Hyperlink">, 2005</span></a>). Cold chain maintenance is the major concern of live attenuated vaccines in tropical countries. Attempts were made to develop thermal adapted vaccine by growing the virus at high temperature (&gt;40°C) (<a href="#Balamurugan-V-2014b"><span class="Hyperlink">Balamurugan et al., 2014b</span></a>; <a href="#Riyesh-T--Balamurugan-V--S"><span class="Hyperlink">Riyesh et al., 2011</span></a>). Being a live attenuated vaccine, animals mount strong immune response against all the viral proteins and therefore differentiating the infected from the vaccinated animals (DIVA) would be difficult. Developing genetically marked vaccines would be advantageous during the eradication phase. Subunit vaccines prepared by expressing the PPRV F and/or H gene in pox and adeno viral vectors found to be experimentally protective (<a href="#Caufour-P--Rufael-T--Lamien-CE--Lancelot-R--Kidane-M-"><span class="Hyperlink">Caufour et al., 2014</span></a>; <a href="#Chandran-D--Reddy-KB--Vijayan-SP--Sugumar-P--Rani-GS--et-al.--2010"><span class="Hyperlink">Chandran et al., 2010</span></a>; <a href="#Chen-W--Hu-S--Qu-L--Hu-Q--Zhang-Q--et-a"><span class="Hyperlink">Chen et al., 2010</span></a>; <a href="#Diallo-A--Mine--Le-2007"><span class="Hyperlink">Diallo et al., 200</span></a><span class="Hyperlink">7</span>; <a href="#Herbert-R--Baron-J--Batten-C--Baron-M--Taylor-G--2014-"><span class="Hyperlink">Herbert et al., 2014</span></a>; <a href="#Rojas-JM--More"><span class="Hyperlink">Rojas et al., 2014</span></a>). Attempt was made to develop a subunit vaccine by expressing the H protein of PPRV in peanut plants (<span class="CharOverride-8">Arachis hypogea</span>) and found to elicit neutralizing antibody responses in sheep (<a href="#Khandelwal-A--Renukaradhya-GJ-"><span class="Hyperlink">Khandelwal et al., 2011</span></a>). The major disadvantage of this strategy is need of multiple doses for protective immunity. Another approach is to manipulate a specific region or epitope of a viral protein of the existing vaccine to obtain positively and or negatively marked vaccines. Recently, two groups succeeded in developing DIVA vaccines using Nigeria 75/1 strain (<a href="#Hu-Q--Chen-W--Huang-K--Baron-MD--Bu-Z--2012"><span class="Hyperlink">Hu et al., 2012</span></a>; <a href="#Muniraju-M--Mahapatra-M--Buczkowski"><span class="Hyperlink">Muniraju et al., 2015</span></a>). Our lab is currently working on development of a DIVA vaccine for PPRV Sungri/96. Experimental combined vaccines against PPR/goatpox (<a href="#Hosamani-M--Singh-SK--Mondal-B--Sen-A--Bhanuprakash-V--et-al.--2006"><span class="Hyperlink">Hosamani et al., 2006</span></a>), and PPR/sheeppox (<a href="#Chaudhary-SS--Pandey-KD--Singh-RP--Verma-PC--Gupta-PK--2009"><span class="Hyperlink">Chaudhary et al., 2009</span></a>) have been shown as promising candidates. Appropriate period of vaccination in kids was found to be four months (<a href="#Balamurugan-V--2012b"><span class="Hyperlink">Balamurugan et al., 2012b</span></a>). </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1"><span class="CharOverride-13">Diagnostics of PPR</span></p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Laboratory confirmation of the disease is usually done through virus isolation and virus neutralization assay, which are time consuming and laborious. Several assays have been described to detect virus-specific antibodies or viral antigens. </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-2--History-in-MM-">Detection of Antibodies Against PPRV </p>
			<p class="Body-Text ParaOverride-1">A monoclonal antibody based competitive ELISA (cELISA) for detection of antibodies against PPRV has been developed in our laboratory (<a href="#Singh-RP--Saravanan-2004a"><span class="Hyperlink">Singh et al., 2004a</span></a>). The assay was compared with VNT and found to be 92.2% and 98.84%, of specificity and sensitivity, respectively. Although, this test is suitable for mass screening, it uses live virus as positive antigen, which could be a disadvantage for use in PPR free countries/regions. To overcome this, we are currently working on a recombinant antigen based competitive ELISA. Similarly, a polyclonal antibody based indirect ELISA was developed for detection of antibodies against PPRV in the serum samples (<a href="#Balamurugan-V-2007"><span class="Hyperlink">Balamurugan et al., 2007</span></a>). The performance of the assay was comparable with that of c-ELISAs. </p>
			<p class="Heading-2--History-in-MM-">&nbsp;</p>
			<p class="Heading-2--History-in-MM-">Detection of PPRV Antigen </p>
		  <p class="Heading-2--History-in-MM-">&nbsp;</p>
			<p class="Heading-3">Virus Isolation </p>
			<p class="Body-Text ParaOverride-1">Virus isolation remains the “<span class="CharOverride-8">gold standard</span>” for the diagnosis of PPR. Blood collected at the height of the temperature is the best material for this purpose. The nasal or ocular swabs or 10% tissue suspension can also be used. The PPRV can be propagated <span class="CharOverride-8">in vitro</span> in several primary bovine and sheep cells, as well as established cell lines like Vero (African green monkey kidney cells) and B95a (Marmoset B-lymphoblastoid cells) (<a href="#Sreenivasa-BP--Singh-RP--Mondal-B--Dhar-P-"><span class="Hyperlink">Sreenivasa et al., 2006</span></a>). In most of these cells, the PPRV manifests morbillivirus-specific CPE by 3-5 days post infection. In some cases, up to five blind passages are needed for isolating the virus. </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-3">Sandwich ELISA</p>
			<p class="Body-Text ParaOverride-1">The sandwich-ELISA for the detection of PPRV antigen in the clinical samples was developed at our laboratory. This assay uses a monoclonal antibody directed against N protein. The test is compared with IC-ELISA (BDSL) and found to have 89% and 93%, sensitivity and specificity, respectively (<a href="#Singh-RP--Sreeniva-2004b"><span class="Hyperlink">Singh et al., 2004b</span></a>). This assay is easy to perform and is routinely adopted by many diagnostic laboratories. Like c-ELISA, in this assay also live attenuated PPRV used as positive antigen. Attempts were made to develop a recombinant N protein based ELISA (<a href="#Yadav-V--Balamurugan-V--Bhanuprakash-V--Sen-A--Bhanot-V--et-al.--2009"><span class="Hyperlink">Yadav et al., 2009</span></a>).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-2--History-in-MM-">Detection of PPRV Nucleic Acid</p>
			<p class="Body-Text ParaOverride-1">Reverse Transcription-Polymerase Chain Reaction (RT-PCR) is the method of choice for detecting nucleic acids of PPRV in clinical samples. RT-PCRs have been reported for detection and differential diagnosis of RP and PPR viruses targeting the N and F gene (<a href="#Balamurugan-V--2006"><span class="Hyperlink">Balamurugan et al., 2006</span></a>; <a href="#Couacy-Hymann-E--Roge"><span class="Hyperlink">Couacy-Hymann et al., 2002</span></a>; <a href="#Forsyth-MA--Barrett-T--1995"><span class="Hyperlink">Forsyth and Barrett, 1995</span></a>). N-Gene based PCR-ELISA has developed at IVRI, Mukteswar to detect PPRV from clinical samples. The test can detect viral RNA in infected tissue culture fluid with a titre as low as 0.01 TCID<span class="CharOverride-14">50 </span>and is used to evaluate only critical samples (<a href="#Saravanan-P--Balamurugan-2007"><span class="Hyperlink">Saravanan et al., 2004</span></a>). PCR based on other genes also are available (<a href="#Brindha-K--Raj-GD--Ganesan-PI--Thiagarajan-V--Nainar-AM--Nachimuthu-K--2001"><span class="Hyperlink">Brindha et al., 2001</span></a>; <a href="#George-A--Dhar-P--Sreenivasa-BP--Singh"><span class="Hyperlink">George et al., 2006</span></a>).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp; </p>
			<p class="Heading-2--History-in-MM-">Real-time RT-PCR and Lateral Flow Assays</p>
			<p class="Body-Text ParaOverride-1">Real time RT- PCR has been used for quantification and diagnosis of PPR virus. M gene-based hydrolysis probe (TaqMan), SYBR Green I based real-time RT-PCR assays targeting the M gene of PPRV are in use (<a href="#Abera-T--Thangavelu-A--2014"><span class="Hyperlink">Abera and Thangavelu, 2014</span></a>; <a href="#Balamurugan-V-2012c"><span class="Hyperlink">Balamurugan et al., 2012c</span></a>). Recently, a novel non-amplification technique was developed to detect nucleic acids of PPRV in which two probes complementary to the target sequences - one conjugated to magnetic microparticles and the second to gold nanoparticles labelled with horseradish peroxidase (<a href="#Tao-C--Li-G--Wang-Y--Huang-H--2013"><span class="Hyperlink">Tao et al., 2013</span></a>). The assay has great potential due to quick performance (45 min) and not requiring expensive instrumentations etc. </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text">Recently, an immunochromatographic test has been developed for the diagnosis PPR under field conditions. The assay has been validated with clinical samples collected in Ivory Coast, Pakistan, Ethiopia and Uganda (<a href="#Baron-J--Fishbourne-E--Couacy-Hyman-E--Abubakar-M--Jones-BA--et-al.--2014"><span class="Hyperlink">Baron et al., 2014</span></a>). </p>
		  <p class="Body-Text">&nbsp;</p>
			<p class="Heading-2--History-in-MM- ParaOverride-1">Loop Mediated Isothermal Amplification (LAMP) Assay </p>
			<p class="Body-Text">LAMP assay for rapid detection of PPRV from clinical samples has been developed using M gene (<a href="#Li-L--Bao-J--Wu-X--Wang-Z--Wang-J--et-al.--2010-.-Rapid-detection-of-peste-des-petits-ruminants-viru"><span class="Hyperlink">Li et al., 2010</span></a>). Similar assay has been developed in our laboratory using N gene and the assay is 100 and 1000 times more sensitive than RT-PCR and s-ELISA, respectively (<a href="#Dadas-RC--Muthuchelvan-D--Pandey-AB--Rajak-KK--Su"><span class="Hyperlink">Dadas et al., 2012</span></a>). </p>
		  <p class="Body-Text">&nbsp;</p>
			<p class="Heading-1--Introduction----">Control program in India</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1"><span class="CharOverride-16">Focused vaccinations in high-risk populations and mass vaccination are necessary to achieve 70 to 80% of herd immunity. The control strategies should follow the “bottom-up” approach (farmers to field veterinarians to policy makers) </span>(<a href="#Singh-RP--2011-"><span class="Hyperlink">Singh, 2011</span></a>)<span class="CharOverride-16">. Besides, strengthening of PPR vaccine production and quality control units, disease diagnostic laboratories, infrastructure for field Veterinarians are also key factors. The NCP-PPR control programme started in 2010 and currently expanded to all the provinces of the country. </span></p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">Conflict of interest</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">There exists no conflict of interest.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">Acknowledgements</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">The authors thank ICAR-Indian Veterinary Research Institute for necessary support.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">Author’s cONTRIBUTION</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Dhanavelu Muthuchelvan, Dheeraj Choudhary, Kaushal Kishor Rajak, Sakshi Bhadouriya and Paramasivam Saravanan collected the literature. Dhanavelu Muthuchelvan wrote the manuscript. Dhanavelu Muthuchelvan, Kaushal Kishor Rajak and Muthannan Andavar Ramakrishnan edited the manuscript. Awadh Bihari Pandey and Raj Kumar Singh performed the final check.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">References</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			
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