DIAGNOSTIC TEST FOR EQUINE ARTERITIS VIRUS MEDIATED DISEASE

Abstract: This invention relates to recombinant DNA and polypeptides encoded thereby which have use in the provision of vaccines, diagnostic test kits and methods of diagnosis and treatment or prophylaxis for equine arteritis virus (EAV) and EAV mediated disease.

Description: DIAGNOSTIC TEST FOR EQUINE ARTERITIS VIRUS MEDIATED DISEASE The present invention relates to recombinant DNA and polypeptides encoded thereby, having use in provision of antibodies, vaccines, diagnostic test kits and methods of diagnosis and treatment or prophylaxis for equine arteritis virus (EAV) and equine arteritis virus mediated disease. Equine viral arteritis, a disease for which equids are the only reported hosts, has been known for some 40 years and manifests itself with widely varying clinical signs. In its most severe form EAV infection causes abortion and foal death which makes it a potentially significant commercial threat to, inter alia, the horse breeding industry. Early veterinary articles refer to it as epizootic cellulitis, pinkeye or equine influenza. Disease outbreaks are identified infrequently and field isolates of the single stranded RNA virus itself are rare. EAV is transmitted by the respiratory and venereal routes, with a 30-60% carrier state existing in seropositive stallions which persistently shed virus in their semen. Thus the venereal route is a particular cause for concern, as these shedding stallions may infect broodmares at mating. In the light of the potential economic importance of the virus and its stud carrier mediated infection capability there exists a requirement for prophylactic treatment and reliable diagnosis of equine viral arteritis, and rapid identification of equids previously exDosed to the infectious agent. Laboratory tests based on ELISA using whole virus as antigen, virus neutralisation (VN) and complement fixation (CF) formats have been developed (see Chirnside (1992) Br Vet J 148, pp 181) . The known ELISA is relatively insensitive when applied to tissues, eg sera, from horses previously vaccinated for other diseases such as influenza and herpesvirus, while the VN and CF formats have limited temporal sensitivity; the VN test is unable to distinguish between vaccination and natural infection. Vaccination procedures have concentrated on safety and efficacy of whole inactivated virus and attenuated live virus vaccines . The live vaccine can induce short term shedding of virus from the nasopharynx and does not prevent this causing infection of commonly housed animals which have not been so treated. It is not yet known if formalinised vaccines or other inactivated vaccine preparations provide reliable protection. Attempts to provide improvements to both diagnostic tests and vaccines have included studies into panels of antibodies raised against various EAV proteins. A 29K envelope protein (G L ) has been identified as antigenic (Balasuriya et al , (1993) J Gen Virol 74, 2525-2529; Deregt et al , (1994) J Gen Virol 75, 2439-2444) and peptides derived from this protein induce a virus neutralising response in horses (Chirnside et al , (1995 ) J Gen Virol in press) . A previous invention (UK Patent Application No GB 9400656.4) provided isolated peptides of G L that produce a potent neutralising immune response against EAV when administered to animals, particularly horses, and these peptides provided sensitive detection of EAV antibodies when used as binding agents in biding assays formats. Further provided was DNA encoding for these peptides. In the first aspect of the present invention there is provided a method for testing for the presence of antibodies to equine arteritis virus comprising use of a peptide or peptide conjugate of the viral nucleocapsid (N) protein as a specific binding agent. Such test is preferably of ELISA format but may use the peptide or conjugate as immobilised binding agent or labelled secondary binding agent in a so-called sandwich assay. Preferred peptides or peptide conjugates of the invention comprise epitopes present in the amino acid sequence corresponding to amino acids 1 to 110 (SEQ ID No.2) of EAV N, more preferably amino acids 1 to 69 of EAV N (SEQ ID No.3) and, additionally, amino acids 1 to 28 (SEQ ID No.4), or a sequence having at least 90% homology to these sequences. These peptides are antigenic and may be used to provide isolated antibodies produced by an immunological response when a suitable host is exposed to the antigen. In a binding assay where the peptide or peptide conjugate is immobilised this method may conveniently be carried out by use of commercially available assay plates onto which the peptide or conjugate is coated by suitable incubation in the known manner. For the purpose of assay, a sample to be screened for EAV antibodies, eg a serum sample, is typically incubated in contact with the plate, eg in the wells, whereafter any EAV antibody present is identified by exposure to eg an anti-horse IgA, IgM, IgG or other immunoglobulin conjugated to a reporter group. Such repor t er group may be in the form of radiolabel, chemical label or a biological label. A typical biological label is an enzyme or cofactor, eg biotin, and is detected by exposure to ail the reactants necessary for a reporter reaction to occur dependent upon the presence of the reporter group. In the case of biotin the well may be exposed to streptavidin-peroxidase and the substrate o- phenyienediamine dihydrochloride and the absorbance of the plate determined at 490nm. In a further example, an immobilised anti-horse IgA, IgM, IgG or other immunoglobulin antibody raised in another animal, may be used to bind a specific class of horse antibody; the immobilised horse antibody provided may then be exposed to a solution containing labelled peptide or conjugate of the invention whereby the presence of anti- EAV antibody is indicated by assay of the amount of label present. Other assay formats such as competitive assays using either bound or unbound peptide or conjugate will occur to those skilled in the art; these will include simple observation of agglutination between peptide or conjugate and the antibody in a simple dilution test. In a further aspect of the present invention there are provided test kits for use in carrying out the assay of the invention characterised in that they comprise a peptide, peptide-conjugate or antibodies of the invention, together with optional agents and items necessary for performing such assays. Such agents and items may include other binding agents or colour forming agents such as labelled antibodies eg biotinylated anti-horse IgG, horseradish peroxidase, streptavidin peroxidase conjugate and o-phenylenediamine dihydrochloride. It will be realised that the term peptide and peptide conjugate as used herein will encompass oligopeptides, polypeptides and proteins as long as they fulfill the criteria of the invention with regard to the immunological activity and content of epitopic sequences. The term conjugate designates conjugation to any physiologically acceptable entity. The peptides, peptide conjugates and binding assays of the present invention will now be described, by way of example only, by reference to the following sequence listing, figures and examples. SEQUENCE LISTING SEQ ID No.l: is the DNA sequence equivalent to the entire EAV genome minus the first 18 bases and the polyA tail. SEQ ID No.2: is the amino acid sequence of the virus nucleocapsid protein (N) encoded by open reading frame (ORF) 7, and that is fused in-frame to GST in FP70 to express rNl-110. SEQ ID No.3: is the amino acid sequence corresponding to amino acids 1 to 69 of the EAV N protein, and that is fused in-frame to GST in FP71 to express rNl-69. SEQ ID No 4. : is the amino acid sequence corresponding to amino acids 1 to 28 of the EAV N protein, and that is fused in-frame to GST in FP7Fspl to express rNl-28. FIGURES Figure 1 shows an immunoblot of purified fusion proteins (Fps 70, 71, 72, 73 and 7Fspl) and glutathione-S- transferase (Gst) with serum from an individual horse pre- and post-EAV infection. For plasmid and fusion protein derivations see Table 1. Figure 2 shows equine sera ELISA absorbance values to recombinant EAV N proteins . ΞLISA plates were coated with 0.5 μg per well of purified fusion protein (FP) or glutathione-S-transferase (GST) . Sera were tested in two replicate wells to each antigen and the absorbance of each well read at 490nm. The GST absorbance was subtracted from the FP absorbance to derive an EAV-specific value. Each bar represents the mean value from two replicates of each serum. Cut-off points determining ELISA seropositivity for each antigen, calculated from the absorbance values of 8 VN- control sera are shown as a horizontal line on each graph: FP70=0.592; FP71=0.483; FP72=0.294; FP73=0.407; FP7Fspl=0.582. The virus neutralising titre (VN titre) of the 8 sera tested are shown on the x-axis as log 10 VN titres. EXAMPLE 1: Production of peptides and conjugates of the invention and DNA and vectors encoding therefore. cDNA encompassing EAV open reading frame (ORF) 7 (den Boon et al [1991] , J Virol, 65, 2910-2920; de Vries et al , [1992] , J Virol 66, 6294-6303) corresponding to the EAV N protein was cloned into the bacterial vectors pGEX-3X and pGΞX-2T (Table 1) and constructs screened for fusion protein expression using PAGE with cloning confirmed by RE digestion analysis and sequencing over the plasmid/insert junctions. Plasmids are referred to as FPx, the expressed recombinant fusion proteins as rNy-z (where y and z refer to amino acid residue numbers in EAV N) . Affinity purified glutathione-S-transferase (GST) fusion proteins were screened for reactivity in immunoblots with a panel of pre- and post-EAV-infection equine sera. Although horse sera exhibit some background absorbence to GST in immunoblots, post-infection sera bound strongly and specifically to fusion proteins containing amino acids Nl- 110 and Nl-69, and failed to bind fusion proteins containing Nl-28, N70-89 and N90-110 specifically (Figure 1) . EXAMPLE 2: ELISA using EAV nucleocapsid (N) fusion proteins Dynatech Immulon 3 microtitre plates were coated with rNl- 69 or rNl-28 antigen by exposure to lOOμl of 5μg/ml antigen in 0.05M carbonate buffer at pH9.6 (Sigma cat No C3041) at 4°C overnight. Plates were washed three times with phosphate buffered saline (PBS) containing 0.05% Tween 3 20 (PBST) and then blocked with lOOμl PBST containing 5% normal goat serum (Seraiab) (PBSTG) for 1 hour at 37°C. Plates were washed again three times with PBST to render them ready for use. Test sera were diluted 1:100 in PBSTG and lOOμl of this solution added to wells prepared as above and incubated for 90 minutes at 37°C. Plates were washed again three times with PBST and solution prepared by diluting goat anti-horse IgG biotin conjugate (KPL catalogue No 162102) 1:1000 in PBSTG and adding lOOμl to each well before being incubated for 90 minutes at 37°C. Plates were washed three times with PBST and a solution prepared by diluting streptavidin-peroxidase conjugate (KPL catalogue No 143000) 1:1000 in PBSTG and adding lOOμl to each well before incubating at room temperature for 30 minutes. Plates were washed three times with PBST and lOOμl 0- phenylenediamine dihydrochloride (Sigma cat. No P8287) (0.5 mg/ml in 0.05 phosphate citrate buffer, pH5.0, Sigma cat. No. P4922)) added to each well and incubated for 10 minutes at room temperature. 50μl 4M H 2 S0 4 was added to stop the reaction and absorbence read at 490nm. Since horse sera at a 1:100 dilution can bind native GST it is necessary to subtract absorbence readings obtained for sera against GST from the GST-fusion protein absorbence. Each serum is tested in duplicate wells against each antigen. Figure 2 shows the results of 8 VN equine sera in ELISA to different recombinant EAV N proteins. Cut-off points determining ELISA seropositivity for each antigen, calculated from the value of 8VN negative equine sera, are shown as a horizontal line on each graph (rNl-110 = 0.592; rNl-69 = 0.483; rN70-89 = 0.294; rN90-110 = 0.407; rNl-28 = 0.582) . From these results rNl-69 and rNl-28 were identified as suitable antigens for the detection of EAV- specific antibodies in ELISA. EXAMPLE 3: ELISA using rNl-69 and rNl-28 binding agents Panels containing seronegative and virus neutralising sera were t es t ed in ELISA to purified rNl-69 and rNl-28 (Table 2) . In ELISAs a recombinant fusion protein containing residues 1 - 69 or 1-28 discriminated between pre- and post- infection equine sera. In additional ELISA tests screening pre- and post-EAV vaccination samples and including isolate specific sera, the rNl-69 and rNl-28 antigens were able to discriminate between samples pre- and post vaccination with Artervac (commercial inactivated virus vaccine) , even in the absence of vaccination induced neutralising antibody, and detect isolate-specific VN sera as seropositive in ELISA. The mean absorbence rising following vaccination were 1.240±0.690 and 0.495+0.352 for rNl-69 and rNl-28 respectively. Table 1 . Nucleocaps id gene constructs and fusion πrocems Plasmiά Ammo Fusion Fusion ZAV Restriction pGEX vector (F?) acid procem protein c2NA digest restriction residue (rN) size clone digest (N) (kDa) 1 3 ' recessed end filled in with the Klenow ragment of DNA poly erase " Vector derived 1 The negacive nu aer corresponds to additional ammo aciαs cloned into pGEX whic are not encoded by ORF 7 2 see de Vries ec ax 1990, Nuclei Acids Research 18, 3241-3247. 11 Table 2 . Comparison of virus neutralising antibody titres and ΞLISA absorbence values Negative controls 32277 0.126 0.170 32278 0.156 0.310 32279 0.098 0.180 32280 0.09S 0.101 32281 0.11S 0.155 32282 0.11S 0.134 32283 0.161 0.204 32284 0.152 0.222 S Post infection 3.746 .250 322S2 3.6 2.504 0.679 32255 2.475 3.660 ..856 32257 2.625 32253 2.700 3.520 1.182 2.536 0.650 322S9 2.550 1.238 0.314 32260 2.850 2.753 2.024 32261 1.87S 32262 2.475 3.00 0.920 Paired vaccination samples 33745 pre 0 - 0.132 - 0.230 post 0.3 - 0.664 + 0.340 33746 pre 0 - 0.353 * 0.344 post 0.45 - 0.967 + 0.580 33747 pre 0 - 0.168 - 0.256 post 0.525 - 2.427 + 1.212 33962 pre 0 - 0.157 - 0.170 post 0 - 0.884 + 0.387 339 63 pre 0 - 0.117 - 0.145 post 0.9 - 2.144 + 0.997 3 3964 pre 0 - 0.156 - 0.175 post 1.572 0.851 33435 pre 0.348 0.286 post 0.3 1.452 0.491 35097 post 1.5 3.226 2.026 35098 post 1.5 3.441 ..908 - 14 - Isolate specific Bucyrus 3.1 - 3.249 - 1.130 84-KY-A1 2.5 - 0.888 - 0.288 roclaw-2 2.2 * 0.424 * 0.276 Arvac 2.5 * 0.422 - 0.319 Killed Bucyrus 1 J 9 - 1.117 + 0.620 1 -jog 10 VN antibody ticre ≥ 0.6 is deemed seropcsicive in Che EAV VN neutralising test b the cu off value to determine seroposicive status was taken as (mean +2SD) of the 8 VN negacive control sera (posicive ≥ 0.177) che cue off value co determine seroposicive stacus was C aken as (mean + 2SD) of che 8 VN negacive concroi sera (posicive ≥ 0.308) pre = Dre-vaccination serum sample DOSC = Dosc-vaccinacion serum sample SEQUENCE LISTING (1) GENERAL INFORMATION: (i) APPLICANT: (A) NAME: The Animal Health Trusc (B) STREET: ?0 Eox 5 (C) CITY: Newmarket (D) STATE: Suffolk (E) COUNTRY: United Kingdom (F) POSTAL CODE (ZIP) : CB8 7DW ( ii ) TITLE OF INVENTION: Diagnostic Test For Equine Arteritis Virus Mediated Disease (iii) NUMBER OF SEQUENCES: 4 (iv) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS (D) SOFTWARE: Patentln Release #1.0, Version #1.30 (EPO) (2) INFORMATION FOR SEQ ID NO: 1: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 12687 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS : single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (Xi) SEQUENCE DESCRIPTION: SEQ ID NO: 1: TGCCATATAC GGCTCACCAC CATATACACT GCAAGAATTA CTATTCTTGT GGGCCCCTCT 60 CGGTAAATCC TAGAGGGCTT TCCTCTCGTT ATTGCGAGAT TCGTCGTTAG ATAACGGCAA 120 GTTCCCTTTC TTACTATCCT ATTTTCATCT TGTGGCTTGA CGGGTCACTG CCATCGTCGT 180 CGATCTCTAT CAACTACCCT TGCGACTATG GCAACCTTCT CCGCTACTGG ATTTGGAGGG 240 AGTTTTGTTA GGGACTGGTC CCTGGACTTA CCCGACGCTT GTGAGCATGG CGCGGGATTG 300 TGCTGCGAAG TGGACGGCTC CACCTTATGC GCCGAGTGTT TTCGCGGTTG CGAAGGAATG 360 GAGCAATGTC CTGGCTTGTT CATGGGACTG TTAAAACTGG CTTCGCCAGT TCCAGTGGGA 420 CATAAGTTCC TGATTGGTTG GTATCGAGCT GCCAAAGTCA CCGGGCGTTA CAATTTCCTT 480 GAGCTGTTGC AACACCCTGC TTTCGCCCAG CTGCGTGTGG TTGATGCTAG GTTAGCCATT 540 GAAGAGGCAA GTGTGTTTAT TTCCACTGAC CACGCGTCTG CTAAGCGTTT CCCTGGCGCT 600 AGATTTGCGC TGACACCGGT GTATGCTAAC GCTTGGGTTG TGAGCCCGGC TGCTAACAGT 660 TTGATAGTGA CCACTGACCA GGAACAAGAT GGGTTCTGCT GGTTAAAACT TTTGCCACCT 720 GACCGCCGTG AGGCTGGTTT GCGGTTGTAT TACAACCATT ACCGCGAACA AAGGACCGGG 780 TGGCTGTCTA AAACAGGACT TCG TTATGG CTTGGAGACC TGGGTTTGGG CATCAATGCG 840 AGCTCTGGAG GGCTGAAATT CCACATTATG AGGGGTTCGC CTCAGCGAGC TTGGCATATC 900 ACAACACGCA GCTGCAAGCT GAAGAGCTAC TACGTTTGTG ACATCTCTGA AGCAGACTGG 960 TCCTGTTTGC CTGCTGGCAA CTACGGCGGC TACAATCCAC CAGGGGACGG AGCTTGCGGT 1020 TACAGGTGCT TGGCCTTCAT GAATGGCGCC ACTGTTGTGT CGGCTGGTTG CAGTTCTGAC 1080 TTGTGGTGTG ATGATGAGTT GGCTTATCGA GTCTTTCAAT TGTCACCCAC GTTCACGGTT 1140 ACCATCCCAG GTGGGCGAGT TTGTCCGAAT GCCAAGTACG CAATGATTTG TGACAAGCAG 1200 CACTGGCGCG TCAAACGTGC AAAGGGCGTC GGCCTGTGTC TCGATGAAAG CTGTTTCAGG 1260 GGCATCTGCA ATTGCCAACG CATGAGTGGA CCACCACCTG CACCCGTGTC AGCCGCCGTG 1320 TTAGATCACA TACTGGAGGC GGCGACGTTT GGCAACGTTC GCGTGGTTAC ACCTGAAGGG 1380 CAGCCACGCC CCGTACCAGC GCCGCGAGTT CGTCCCAGCG CCAACTCTTC TGGAGATGTC 1440 AAAGATCCGG CGCCCGTTCC GCCAGTACCA AAACCAAGGA CCAAGCTTGC CACACCGAAC 1500 CCAACTCAGG CGCCCATCCC AGCACCGCGC ACGCGACTTC AAGGGGCCTC AACACAGGAG 1560 CCACTGGCGA GTGCAGGAGT TGCTTCTGAC TCGGCACCTA AATGGCGTGT GGCCAAAACT 1620 GTGTACAGCT CCGCGGAGCG CTTTCGGACC GAACTGGTAC AACGTGCTCG GTCCGTTGGG 1680 GACGTTCTTG TTCAAGCGCT ACCGCTCAAA ACCCCAGCAG TGCAGCGGTA TACCATGACT 1740 CTGAAGATGA TGCGTTCACG CTTCAGTTGG CACTGCGACG TGTGGTACCC TTTGGCTGTA 1800 ATCGCTTGTT TGCTCCCTAT ATGGCCATCT CTTGCTTTGC TCCTTAGCTT TGCCATTGGG 1860 TTGATACCCA GTGTGGGCAA TAATGTTGTT CTGACAGCGC TTCTGGTTTC ATCAGCTAAT 1920 TATGTTGCGT CAATGGACCA TCAATGTGAA GGTGCGGCTT GCTTAGCCTT GCTGGAAGAA 1980 GAACACTATT ATAGAGCGGT CCGTTGGCGC CCGATTACAG GCGCGCTGTC GCTTGTGCTC 2040 AATTTACTGG GGCAGGTAGG CTATGTAGCT CGTTCCACCT TTGATGCAGC TTATGTTCCT 2100 TGCACTGTGT TCGATCTTTG CAGCTTTGCT ATTCTGTACC TCTGCCGCAA TCGTTGCTGG 2160 AGATGCTTCG GACGCTGTGT GCGAGTTGGG CCTGCCACGC ATGTTTTGGG CTCCACCGGG 2220 CAACGAGTTT CCAAACTGGC GCTCATTGAT TTGTGTGACC ACTTTTCAAA GCCCACCATC 2280 GATGTTGTGG GCATGGCAAC TGGTTGGAGC GGATGTTACA CAGGAACCGC CGCAATGGAG 2340 CGTCAGTGTG CCTCTACGGT GGACCCTCAC TCGTTCGACC AGAAGAAGGC AGGAGCGACT 2400 GTTTACCTCA CCCCCCCTGT CAACAGCGGG TCAGCGCTGC AGTGCCTCAA TGTCATGTGG 2460 AAGCGACCAA TTGGGTCCAC TGTCCTTGGG GAACAAACAG GAGCTGTTGT GACGGCGGTC 2520 AAGAGTATCT CTTTCTCACC TCCCTGCTGC GTCTCTACCA CTTTGCCCAC CCGACCCGGT 2580 GTGACCGTTG TCGACCATGC TCTTTACAAC CGGTTGACTG CTTCAGGGGT CGATCCCGCT 2640 TTATTGCGTG TTGGGCAAGG TGATTTTCTA AAACTTAATC CGGGGTTCCG GCTGATAGGT 2700 GGATGGATTT ATGGGATATG CTATTTTGTG TTGGTGGTTG TGTCAACTTT TACCTGCTTA 2760 CCTATCAAAT GTGGCATTGG CACCCGCGAC CCTTTCTGCC GCAGAGTGTT TTCTGTACCC 2820 GTCACCAAGA CCCAAGAGCA CTGCCATGCT GGAATGTGTG CTAGCGCTGA AGGCATCTCT 2880 CTGGACTCTC TGGGGTTAAC TCAGTTACAA AGTTACTGGA TCGCAGCCGT CACTAGCGGA 2940 TTAGTGATCT TGTTGGTCTG CCACCGCCTG GCCATCAGCG CCTTGGACTT GTTGACTCTA 3000 GCTTCCCCTT TAGTGTTGCT TGTGTTCCCT TGGGCATCTG TGGGGCTTTT ACTTGCTTGC 3060 AGTCTCGCTG GTGCTGCTGT GAAAATACAG TTGTTGGCGA CGCTTTTTGT GAATCTGTTC 3120 TTTCCCCAAG CTACCCTTGT CACTATGGGA TACTGGGCGT GCGTGGCGGC TTTGGCCGTT 3180 TACAGTTTGA TGGGCTTGCG AGTGAAAGTG AATGTGCCCA TGTGTGTGAC ACCTGCCCAT 3240 TTTCTGCTGC TGGCGAGGTC AGCTGGACAG TCAAGAGAGC AGATGCTCCG GGTCAGCGCT 3300 GCTGCCCCCA CCAATTCACT GCTTGGAGTG GCTCGTGATT GTTATGTCAC AGGCACAACT 3360 CGGCTGTACA TACCCAAGGA AGGCGGGATG GTGTTTGAAG GGCTATTCSG GTCACCGAAG 3420 GCGCGCGGCA ACGTCGGCTT CGTGGCTGGT AGCAGCTACG GCACAGGGTC AGTGTGGACC 3480 AGGAACAACG AGGTCGTCGT ACTGACAGCG TCACACGTGG TTGGCCGCGC TAACATGGCC 3540 ACTCTGAAGA TCGGTGACGC AATGCTGACT CTGACTTTCA AAAAGAATGG CGACTTCGCC 3600 GAGGCAGTGA CGACACAGTC CGAGCTCCCA GGCAATTGGC CACAGTTGCA TTTCGCCCAA 3660 CCAACAACCG GGCCCGCTTC ATGGTGCACT GCCACAGGAG ATGAAGAAGG CTTGCTCAGT 3720 GGCGAGGTTT GTCTGGCGTG GACTACTAGT GGCGACTCTG GATCTGCAGT GGTTCAGGGT 3780 GACGCTGTGG TAGGGGTCCA CACCGGTTCG AACACAAGTG GTGTTGCCTA CGTGACCACC 3840 CCAAGCGGAA AACTCCTTGG CGCCGACACC GTGACTTTGT CATCACTGTC AAAGCATTTC 3900 ACAGGCCCTT TGACATCAAT CCCGAAGGAC ATCCCTGACA ACATTATTGC CGATGTTGAT 3960 GCTGTTCCTC GTTCTCTGGC CATGCTGATT GATGGCTTAT CCAATAGAGA GAGCAGCCTT 4020 CTGGACCTC AGTTGTTGTT AATTGCTTGT TTTATGTGGT CTTATCTTAA CCAACCTGCT 4080 TACTTGCCTT ATGTGCTGGG CTTCTTTGCC GCTAACTTCT TCCTGCCAAA AAGTGTTGGC 4140 CGCCCTGTGG TCACTGGGCT TCTATGGTTG TGCTGCCTCT TCACACCGCT TTCCATGCGC 4200 TTGTGCTTGT TCCATCTGGT CTGTGCTACC GTCACGGGAA ACGTGATATC TTTGTGGTTC 4260 TACATCACTG CCGCTGGCAC GTCTTACCTT TCTGAGATGT GGTTCGGAGG CTATCCCACC 4320 ATGTTGTTTG TGCCACGGTT CCTAGTGTAC CAGTTCCCCG GCTGGGCTAT TGGCACAGTA 4380 CTAGCGGTAT GCAGCATCAC CATGCTGGCT GCTGCCCTCG GTCACACCCT GTTACTGGAT 4440 GTGTTCTCCG CCTCAGGTCG CTTTGACAGG ACTTTCATGA TGAAATACTT CCTGGAGGGA 4500 GGAGTGAAAG AGAGTGTCAC CGCCTCAGTC ACCCGCGCTT ATGGCAAACC AATTACCCAG 4560 GAGAGTCTCA CTGCAACATT AGCTGCCCTC ACTGATGATG ACTTCCAATT CCTCTCTGAT 4620 GTGCTTGACT GTCGGGCCGT CCGATCGGCA ATGAATCTCG GTGCCGCTCT CACAAGTTTT 4680 CAAGTGGCGC AGTATCGTAA CATCCTTAAT GCATCCTTGC AAGTCGATCG TGACGCTGCT 4740 CGTAGTCGCA GACTAATGGC AAAACTGGCT GATTTTGCGG TTGAACAAGA AGTAACAGCT 4800 GGAGACCGTG TTGTGGTTAT CGACGGTCTG GACCGCATGG CTCACTTCAA AGACGATTTG 4860 GTGCTGGTTC CTTTGACCAC CAAAGTAGTA GGCGGTTCTA GGTGCACCAT TTGTGACGTC 4920 GTTAAGGAAG AAGCCAATGA CACCCCAGTT AAGCCAATGC CCAGCAGGAG ACGCCGCAAG 4980 GGCCTGCCTA AAGGTGCTCA GTTGGAGTGG GACCGTCACC AGGAAGAGAA GAGGAACGCC 5040 GGTGATGATG ATTTTGCGGT CTCGAATGAT TATGTCAAGA GAGTGCCAAA GTACTGGGAT 5100 CCCAGCGACA CCCGAGGCAC GACAGTGAAA ATCGCCGGCA CTACCTATCA GAAAGTGGTT 5160 GACTATTCAG GCAATGTGCA TTACGTGGAG CATCAGGAAG ATCTGCTAGA CTACGTGCTG 5220 GGCAAGGGGA GCTATGAAGG CCTAGATCAG GACAAAGTGT TGGACCTCAC AAACATGCTT 5280 AAAGTGGACC CCACGGAGCT CTCCTCCAAA GACAAAGCCA AGGCGCGTCA CGTTGCTCAT 5340 CTGCTGTTGG ATCTGGCTAA CCCAGTTGAG GCAGTGAATC AGTTAAACTG AGAGCGCCCC 5400 ACATCTTTCC CGGCGATGTG GGGCGTCGGA CCTTTGCTGA CTCTAAAGAC AAGGGTTTCG 5460 TGGCTCTACA CAGTCGCACA ATGTTTTTAG CTGCCCGGGA CTTTTTATTT AACATCAAAT 5520 TTGTGTGCGA CGAAGAGTTC ACAAAGACCC CAAAAGACAC ACTGCTTGGG TACGTACGCG 5580 CCTGCCCTGG TTACTGGTTT ATTTTCCGTC GTACGCACCG GTCGCTGATT GATGCATACT 5640 GGGACAGTAT GGAGTGCGTT TACGCGCTTC CCACCATATC TGATTTTGAT GTGAGCCCAG 5700 GTGACGTCGC AGTGACGGGC GAGCGATGGG ATTTTGAATC TCCCGGAGGA GGCCGTGCAA 5760 AACGTCTCAC AGCTGATCTG GTGCACGCTT TTCAAGGGTT CCACGGAGCC TCTTATTCCT 5820 ATGATGACAA GGTGGCAGCT GCTGTCAGTG GTGACCCGTA TCGGTCGGAC GGCGTCTTGT 5880 ATAACACCCG TTGGGGCAAC ATTCCATATT CTGTCCCAAC CAATGCTTTG GAAGCCACAG 5940 CTTGCTACCG TGCTGGATGT GAGGCCGTTA CCGACGGGAC CAACGTCATC GCAACAATTG 6000 GGCCCTTCCC GGAGCAACAA CCCATACCGG ACATCCCAAA GAGCGTGCTT GACAACTGCG 6060 CTGACATCAG CTGTGACGCT TTCATAGCGC CCGCTGCAGA GACAGCCCTG TGTGGAGATT 6120 TAGAGAAATA CAACCTATCC ACGCAGGGTT TTGTGTTGCC TAGTGTTTTC TCCATGGTGC 6180 GGGCGTACTT AAAAGAGGAG ATTGGAGACG CTCCACCACT CTACTTGCCA TCTACTGTAC 6240 CATCTAAAAA TTCACAAGCC GGAATTAACG GCGCTGAGTT TCCTACAAAG TCTTTACAGA 6300 GCTACTGTTT GATTGATGAC ATGGTGTCAC AGTCCATGAA AAGCAATCTA CAAACCGCCA 6360 CCATGGCGAC TTGTAAACGG CAATACTGTT CCAAATACAA GATTAGGAGC ATTCTGGGCA 6420 CCAACAATTA CATTGGCCTA GGTTTGCGTG CCTGCCTTTC GGGGGTTACG GCCGCATTCC 6480 AAAAAGCTGG AAAGGATGGG TCACCGATTT ATTTGGGCAA GTCAAAATTC GACCCGATAC 6540 CAGCTCCTGA CAAGTACTGC CTTGAAACAG ACCTGGAGAG TTGTGATCGC TCCACCCCGG 6600 CTTTGGTGCG TTGGTTCGCT ACTAATCTTA TTTTTGAGCT AGCTGGCCAG CCCGAGTTGG 6660 TGCACAGCTA CGTGTTGAAT TGCTGTCACG ATCTAGTTGT GGCGGGTAGT GTAGCATTCA 6720 CCAAACGCGG GGGTTTGTCA TCTGGAGACC CTATCACTTC CATTTCCAAT ACCATCTATT 6780 CATTGGTGCT GTACACCCAG CACATGTTGC TATGTGGACT TGAAGGCTAT TTCCCAGAGA 6840 TTGCAGAAAA ATATCTTGAT GGCAGCCTGG AGCTGCGGGA CATGTTCAAG TACGTTCGAG 6900 TGTACATCTA CTCGGACGAT GTGGTTCTAA CCACACCCAA CCAGCATTAC GCGGCCAGCT 6960 TTGACCGCTG GGTCCCCCAC CTGCAGGCGC TGCTAGGTTT CAAGGTTGAC CCAAAGAAAA 7020 CTGTGAACAC CAGCTCCCCT TCCTTTTTGG GCTGCCGGTT CAAGCAAGTG GACGGCAAGT 7080 GTTATCTAGC CAGTCTTCAG GACCGCGTTA CACGCTCTCT GTTATACCAC ATTGGTGCAA 7140 AGAATCCCTC AGAGTACTAT GAAGCTGCTG TTTCCATCTT TAAGGACTCC ATTATCTGCT 7200 GTGATGAAGA CTGGTGGACG GACCTCCATC GACGTATCAG TGGCGCTGCG CGTACCGACG 7260 GAGTTGAGTT CCCCACCATT GAAATGTTAA CATCCTTCCG CACCAAGCAG TATGAGAGTG 7320 CCGTGTGCAC AGTTTGTGGG GCCGCCCCCG TGGCCAAGTC TGCTTGTGGA GGGTGGTTCT 7380 GTGGCAATTG TGTCCCGTAC CACGCGGGTC ATTGTCACAC AACCTCGCTC TTCGCCAACT 7440 GCGGGCACGA CATCATGTAC CGCTCCACTT ACTGCACAAT GTGTGAGGGT TCCCCAAAAC 7500 AGATGGTACC AAAAGTGCCT CACCCGATCC TGGATCATTT GCTGTGCCAC ATTGATTACG 7560 GCAGTAAAGA GGAACTAACT CTGGTAGTGG CGGATGGTCG AACAACATCA CCGCCCGGGC 7620 GCTACAAAGT GGGTCACAAG GTAGTCGCCG TGGTTGCAGA TGTGGGAGGC AACATTGTGT 7680 TTGGGTGCGG TCCTGGATCA CACATCGCAG TACCACTTCA GGATACGCTC AAGGGCGTGG 7740 TGGTGAATAA AGCTCTGAAG AACGCCGCCG CCTCTGAGTA CGTGGAAGGA CCCCCTGGGA 7800 GTGGGAAGAC TTTTCACCTG GTCAAAGATG TGCTAGCCGT GGTCGGTAGC GCGACCTTGG 7860 TTGTGCCCAC CCACGCGTCC ATGCTGGACT GCATCAACAA GCTCAAACAA GCGGGCGCCG 7920 ATCCATACTT TGTGGTGCCC AAGTATACAG TTCTTGACTT TCCCCGGCCT GGCAGTGGAA 7980 ACATCACAGT GCGACTGCCA CAGGTCGGAA CCAGTGAGGG AGAAACCTTT GTGGATGAGG 8040 TGGCCTACTT CTCACCAGTG GATCTGGCGC GCATTTTAAC CCAGGGTCGA GTCAAGGGTT 8100 ACGGTGATTT AAATCAGCTC GGGTGCGTCG GACCCGCGAG CGTGCCACGT AACCTTTGGC 8160 TCCGACATTT TGTCAGCCTG GAGCCCTTGC GAGTGTGCCA TCGATTCGGC GCTGCTGTGT 8220 GTGATTTGAT CAAGGGCATT TATCCTTATT ATGAGCCAGC TCCACATACC ACTAAAGTGG 8280 TGTTTGTGCC AAATCCAGAC TTTGAGAAAG GTGTAGTCAT CACCGCCTAC CACAAAGATC 8340 GCGGTCTTGG TCACCGCACA ATTGATTCAA TTCAAGGCTG TACATTCCCT GTTGTGACTC 8400 TTCGACTGCC CACACCCCAA TCACTGACGC GCCCGCGCGC AGTTGTGGCG GTTACTAGGG 8460 CGTCTCAGGA ATTATACATC TACGACCCCT TTGATCAGCT TAGCGGGTTG TTGAAGTTCA 8520 CCAAGGAAGC AGAGGCGCAG GACTTGATCC ATGGCCCACC TACAGCATGC CACCTGGGCC 8580 AAGAAATTGA CCTTTGGTCC AATGAGGGCC TCGAATATTA CAAGGAAGTC AACCTGCTGT 8640 ACACACACGT CCCCATCAAG GATGGTGTAA TACACAGTTA CCCTAATTGT GGCCCTGCCT 8700 GTGGCTGGGA AAAGCAATCC AACAAAATTT CGTGCCTCCC GAGAGTGGCA CAAAATTTGG 8760 GCTACCACTA TTCCCCAGAC TTACCAGGAT TTTGCCCCAT ACCAAAAGAA CTCGCTGAGC 8820 ATTGGCCCGT AGTGTCCAAT GATAGATACC CGAATTGCTT GCAAATTACC TTACAGCAAG 8880 TATGTGAACT CAGTAAACCG TGCTCAGCGG GCTATATGGT TGGACAATCT GTTTTCGTGC 8940 AGACGCCTGG TGTGACATCT TACTGGCTTA CTGAATGGGT CGACGGCAAA GCGCGTGCTC 9000 TACCAGATTC CTTATTCTCG TCCGGTAGGT TCGAGACTAA CAGCCGCGCT TCCTCGATG 9060 AAGCCGAGGA AAAGTTTGCC GCCGCTCACC CTCATGCCTG TTTGGGAGAA ATTAATAAGT 9120 CCACCGTGGG AGGATCCCAC TTCATCTTTT CCCAATATTT ACCACCATTG CTACCCGCAG 9180 ACGCTGTTGC CCTGGTAGGT GCTTCATTGG CTGGGAAAGC TGCTAAAGCT GCTTGCAGCG 9240 TTGTTGATGT CTATGCTCCA TCATTTGAAC CTTATCTACA CCCTGAGACA CTGAGTCGCG 9300 TGTACAAGAT TATGATCGAT TTCAAGCCGT GTAGGCTTAT GGTGTGGAGA AACGCGACCT 9360 TTTATGTCCA AGAGGGTGTT GATGCAGTTA CATCAGCACT AGCAGCTGTG TCCAAACTCA 9420 TCAAAGTGCC GGCCAATGAG CCTGTTTCAT TCCATGTGGC ATCAGGGTAC AGAACCAACG 9480 CGCTGGTAGC GCCCCAGGCT AAAATTTCAA TTGGAGCCTA CGCCGCCGAG TGGGCACTGT 9540 CAACTGAACC GCCACCTGCT GGTTATGCGA TCGTGCGGCG ATATATTGTA AAGAGGCTCC 9600 TCAGCTCAAC AGAAGTGTTC TTGTGCCGCA GGGGTGTTGT GTCTTCCACC TCAGTGCAGA 9660 CCATTTGTGC ACTAGAGGGA TGTAAACCTC TGTTCAACTT CTTACAAATT GGTTCAGTCA 9720 TTGGGCCCGT GTGATGGGCT TAGTGTGGTC ACTGATTTCA AATTCTATTC AGACTATTAT 9780 TGCTGATTTT GCTATTTCTG TGATTGATGC AGCGCTTTTC TTTCTCATGC TACTTGCATT 9840 GGCTGTTGTT ACTGTGTTTC TTTTCTGGCT CATTGTTGCC ATCGGCCGCA GCTTGGTGGC 9900 GCGGTGTTCA CGAGGTGCGC GTTACAGACC TGTTTAAGGA TTTGCAGTGC GACAACCTGC 9960 GCGCGAAAGA TGCCTTCCCG AGTCTGGGAT ATGCTCTGTC GATTGGCCAG TCGAGGCTAT 10020 CGTATATGCT GCAGGATTGG TTGCTTGCTG CGCACCGCAA GGAAGTTATG CCTTCCAATA 10080 TCATGCCTAT GCCCGGTCTT ACTCCTGATT GCTTTGACCA TCTGGAGTCT TCTAGCTATG 10140 CTCCATTTAT CAATGCCTAT CGGCAGGCAA TTTTGAGTCA ATACCCACAA GAGCTCCAGC 10200 TCGAAGCCAT CAACTGTAAA TTGCTTGCTG TGGTTGCACC GGCATTGTAT CATAATTACC 10260 ATCTAGCCAA TTTGACCGGA CCGGCCACAT GGGTCGTGCC TACAGTGGGC CAGTTGCACT 10320 ATTATGCTTC TTCCTCTATT TTTGCTTCAT CTGTGGAAGT GTTGGCAGCA ATAATACTAC 10380 TATTTGCATG CATACCACTA GTGACACGAG TGTACATCTC TTTTACGCGG CTAATGTCAC 10440 CTTCCCGTCG CACTTCCAGC GGCACTTTGC CGCGGCGCAA GATTTTGTAG TGCACACGGG 10500 TTATGAATAT GCCGGGGTCA CTATGTTAGT GCACTTGTTT GCCAACTTGG TTCTGACATT 10560 TCCGAGCTTA GTTAATTGTT CCCGCCCTGT GAATGTCTTT GCTAATGCTT CTTGCGTGCA 10620 AGTGGTTTGT AGTCATACCA ACTCAACTAC TGGCTTGGGT CAACTTTCTT TTTCCTTTGT 10680 AGATGAAGAT CTACGGCTGC ATATCAGGCC TACTCTTATT TGTTGGTTTG CCTTGTTGTT 10740 GGTGCACTTT CTACCCATGC CACGCTGCAG AGGCTCGTAA TTTTACTTAC ATTAGTCATG 10800 GATTGGGCCA CGTGCACGGT CATGAGGGGT GTAGGAATTT TATTAATGTC ACTCATTCTG 10860 CATTTCTTTA TCTTAATCCC ACCACTCCCA CTGCGCCGGC TATAACTCAT TGTTTACTTC 10920 TGGTTCTGGC AGCCAAAATG GAACACCCAA ACGCTACTAT CTGGCTGCAG CTGCAGCCGT 10980 TTGGGTATCA TGTGGCTGGC GATGTCATTG TCAACTTGGA AGAGGACAAG AGGCATCCTT 11040 ACTTTAAACT TTTGAGAGCG CCGGCTTTAC CGCTTGGTTT TGTGGCTATA GTTTATGTTC 11100 TTTTACGACT GGTACGTTGG GCTCAACGAT GTTATCTATG ATTGTATTGC TATTCTTGCT 11160 TTGGGGTGCG CCATCACATG CTTACTTCTC ATACTACACC GCTCAGCGCT TCACAGACTT 11220 CACCTTGTGT ATGCTGACGG ATCGCGGCGT TATTGCCAAT TTGCTGCGAT ATGATGAGCA 11280 CACTGCTTTG TACAATTGTT CCGCCAGTAA AACCTGTTGG TATTGCACAT TCCTGGACGA 11340 ACAGATTATC ACGTTTGGAA CCGATTGTGA TGACACCTAC GCGGTCCCAG TTGCTGAGGT 11400 CCTGGAACAG GCGCATGGAC CGTACAGTGC GCTGTTTGAT GACATGCCCC CTTTTATTTA 11460 CTATGGCCGT GAATTCGGCA TAGTTGTGTT GGATGTGTTT ATGTTCTATC CCGTTTTAGT 11520 TCTGTTTTTC TTATCAGTAC TACCCTATGC TACGCTTATT CTTGAAATGT GTGTATCTAT 11580 TCTGTTTATA ATCTATGGCA TTTACAGCGG GGCCTACTTG GCCATGGGCA TATTTGCGGC 11640 CACGCTTGCT ATACATTCAA TTGTGGTCCT CCGCCAATTA CTGTGGTTAT GCCTGGCTTG 11700 GCGATACCGC TGTACGCTTC ACGCGTCCTT TATATCAGCT GAGGGGAAAG TGTACCCCGT 11760 AGACCCCGGA CTCCCGGTTG CCGCCGTGGG CAATCGGTTG TTAGTCCCAG GTAGGCCCAC 11820 TATCGATTAT GCAGTGGCCT ACGGCAGCAA AGTCAACCTT GTGAGGTTGG GGGCAGCTGA 11880 GGTATGGGAG CCATAGATTC ATTTTGTGGT GACGGGATTT TAGGTGAGTA TCTAGATTAC 11940 TTTATTCTGT CCGTCCCACT CTTGCTGTTG CTTACTAGGT ATGTAGCATC TGGGTTAGTG 12000 TATGTTTTGA CTGCCTTGTT CTATTCCTTT GTATTAGCAG CTTATATTTG GTTTGTTATA 12060 GTTGGAAGAG CCTTTTCTAC TGCTTATGCT TTTGTGCTTT TGGCTGCTTT TCTGTTATTA 12120 GTAATGAGGA TGATTGTGGG TATGATGCCT CGTCTTCGGT CCATTTTCAA CCATCGCCAA 12180 CTGGTGGTAG CTGATTTTGT GGACACACCT AGTGGACCTG TTCCCATCCC CCGCTCAACT 12240 ACTCAGGTAG TGGTTCGCGG CAACGGGTAC ACCGCAGTTG GTAACAAGCT TGTCGATGGC 12300 GTCAAGACGA TCACGTCCGC AGGCCGCCTC TTTTCGAAAC GGACGGCGGC GACAGCCTAC 12360 AAGCTACAAT GACCTACTGC GCATGTTTGG TCAGATGCGG GTCCGCAAAC CGCCCGCGCA 12420 ACCCACTCAG GCTATTATTG CAGAGCCTGG AGACCTTAGG CATGATTTAA ATCAACAGGA 12480 GCGCGCCACC CTTTCGTCGA ACGTACAACG GTTCTTCATG ATTGGGCATG GTTCACTCAC 12540 TGCAGATGCC GGAGGACTCA CGTACACCGT CAGTTGGGTT CCTACCAAAC AAATCCAGCG 12600 CAAAGTTGCG CCTCCAGCAG GGCCGTAAGA CGTGGATATT CTCCTGTGTG GCGTCATGTT 12660 GAAGTAGTTA TTAGCCACCC AGGAACC 12687 (2) INFORMATION FOR SEQ ID NO: 2: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 110 amino acids (B) TYPE: amino acid (C) STRANDEDNESS : single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2: Met Ala Ser Arg Arg Ser Arg Pro Gin Ala Ala Ser Phe Arg Asn Gly 1 5 10 15 Arg Arg Arg Gin Pro Thr Ser Tyr Asn Asp Leu Leu Arg Met Phe Gly 20 25 30 Gin Met Arg Val Arg Lys Pro Pro Ala Gin Pro Thr Gin Ala lie He 35 40 45 Ala Glu Pro Gly Asp Leu Arg His Asp Leu Asn Gin Gin Glu Arg Ala 50 55 60 Thr Leu Ser Ser Asn Val Gin Arg Phe Phe Met He Gly His Gly Ser 65 70 75 80 Leu Thr Ala Asp Ala Gly Gly Leu Thr Tyr Thr Val Ser Trp Val Pro 85 90 95 Thr Lys Gin He Gin Arg Lys Val Ala Pro Pro Ala Gly Pro 100 05 HO (2) INFORMATION FOR SEQ ID NO: 3: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 69 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide ( i) SEQUENCE DESCRIPTION: SEQ ID NO: 3: Met Ala Ser Arg Arg Ser Arg Pro Gin Ala Ala Ser Phe Arg Asn Gly 1 5 10 15 Arg Arg Arg Gin Pro Thr Ser Tyr Asn Asp Leu Leu Arg Met Phe Gly 20 25 30 Gin Met Arg Val Arg Lys Pro Pro Ala Gin Pro Thr Gin Ala He He 35 40 45 Ala Glu Pro Gly Asp Leu Arg His Asp Leu Asn Gin Gin Glu Arg Ala 50 55 60 Thr Leu Ser Ser Asn 65 (2) INFORMATION FOR SEQ ID NO: 4: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 28 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 4: Met Ala Ser Arg Arg Ser Arg Pro Gin Ala Ala Ser Phe Arg Asn Gly 1 5 10 15 Arg Arg Arg Gin Pro Thr Ser Tyr Asn Asp Leu Leu 20 25

Claims: Cλfriτns 1. An antigenic peptide which includes an epitope included in an amino acid sequence selected from the group which comprises : a) amino acid residues 1 to 110 of EAV N (SEQ ID NO: 2) b) amino acid residues 1 to 69 of EAV N (SEQ ID NO: 3) c) amino acid residues 1 to 28 of EAV N (SEQ ID NO: 4) d) a sequence of amino acid residues having at least 90% homology to any one of sequences (a) , (b) or (c) . 2. A DNA sequence encoding an amino acid sequence selected from the group which comprises : a) amino acid residues 1 to 110 of EAV N (SEQ ID NO: 2) b) amino acid residues 1 to 69 of EAV N (SEQ ID NO: 3) c) amino acid residues 1 to 28 of EAV N (SEQ ID NO: 4) d) a sequence of amino acid residues having at least 90% homology to any one of sequences (a) , (b) or (c) . 3. A conjugate which comprises an antigenic peptide according to claim 1 together with a physiologically acceptable entity. 4. A labelled peptide according to claim 1 or a labelled conjugate according to claim 3 wherein the label is selected from the group consisting of: a radiolabel; a chemical label; a biological label. 5. A vaccine which comprises a pharmaceutical preparation of an antigenic peptide according to claim 1 or a conjugate according to claim 3. 6. A method for testing for the presence of antibodies to equine arteritis virus (EAV) which comprises the step of using a peptide or peptide conjugate of the viral nucleocapsid (N) protein as a specific binding agent. 7. A method according to claim 6 wherein the method uses the peptide or conjugate in an ELISA, or the peptide or peptide conjugate is an immobilised binding agent or labelled secondary binding agent in a sandwich assay. 8. A method according to claim 6'or 7 wherein the peptide according to claim 1 or peptide conjugate according to claim 3 is used. 9. A method according to claim 6 or 7 wherein the labelled peptide or labelled conjugate according to claim 4 is used. 10. An antigenic peptide according to claim 1 or a conjugate according to claim 3 for use as a medicament in the treatment or prophylaxis of equine arteritis virus mediated disease. 11. Use of an antigenic peptide according to claim 1, DNA according to claim 2 or a conjugate according to claim 3 in the manufacture of a medicament for treatment or prophylaxis of equine arteritis virus mediated disease. 12 . A method of treatment or prophylaxis of equine arteritis virus mediated disease which comprises administering a pharmacologically active amount of an an t igenic peptide according to claim 1 or a conjugate according to claim 3. 13. A test kit for use in a method according to claim 8 which kit includes a peptide according to claim 1 or a conjugate according to claim 3. 14. A test kit for use in a method according to claim 9 which kit includes a peptide or a conjugate according to claim 4. 15. An isolated antibody produced by an immunological response to a peptide according to claim 1 or 4 or a peptide conjugate according to claim 3 or 4. 16. A test kit for use in a method for testing for the presence of antibodies to equine arteritis virus (EAV) which kit includes an antibody according to claim 15.

Inventors: CHIRNSIDE EWAN DOUGLAS, CHIRNSIDE, EWAN, DOUGLAS
Applicants: ANIMAL HEALTH TRUST, CHIRNSIDE EWAN DOUGLAS, CHIRNSIDE, EWAN, DOUGLAS
Application year: 1996
Main classification: C12N15
IPC codes: A61K38, A61K39, A61P31, C07K14, C12N15