Method of rapidly producing improved vaccines for animals

Abstract

A method of quickly producing a vaccine for a biotype of pathogenic microorganism is described, where a nucleic acid molecule or fragment thereof is obtained from a biological sample from an animal exposed to the microorganism, a protective molecule is prepared based on the nucleic acid molecule of interest or fragment thereof, and administered to an animal which has been or is as risk of being exposed to the microorganism. A protective response to the biotype of the microorganism is obtained in the animal.

What is claimed is: 
     
         1 . A method of producing a vaccine to protect an animal from a biotype of a microorganism, the method comprising,
 a) identifying a nucleic acid molecule of interest or fragment thereof (NOI) of said microorganism, said NOI known to be capable of producing a protective molecule;   b) obtaining a biological sample from at least one animal where said animal has been exposed to said microorganism;   c) obtaining from said sample said NOI of said microorganism without the need to isolate said microorganism;   d) producing a protective molecule from said NOI, said protective molecule selected from the group consisting of:
 (i) a nucleic acid molecule comprising the sequence of said NOI or a fragment thereof; 
 (ii) a polypeptide or fragment thereof expressed by said NOI; 
 (iii) at least one RNA molecule comprising a sequence that corresponds to all or a portion of said NOI and forms a dsRNA; 
 (iv) at least one RNA molecule that is antisense to all or a portion of said NOI; 
 (v) a nucleic acid molecule producing (iii) or (iv); and 
 (vi) an RNA particle comprising or producing any one of (i)-(v); 
   e) producing a vaccine comprising said protective molecule, wherein said vaccine does not comprise said microorganism or a replicable or living pathogenic microorganism; and   f) providing said protective molecule in said vaccine in an amount that when administered to an animal protects said animal from said biotype of said microorganism.   
     
     
         2 . The method of  claim 1 , wherein said vaccine is produced in one month or less from the time of obtaining said sample. 
     
     
         3 . The method of  claim 1 , wherein said vaccine is produced in seven days or less from the time of obtaining said sample. 
     
     
         4 . The method of  claim 1 , further comprising introducing an alphavirus replicon vector comprising said NOI into a host cell, expressing a polypeptide or fragment thereof from said NOI, and extracting said polypeptide or fragment from said host, wherein said extracted polypeptide or fragment thereof comprises said protective molecule. 
     
     
         5 . The method of  claim 4 , wherein said vector comprises a Venezuelan Equine Encephalitis (VEE) vector, said host comprises Vero cells and said polypeptide or fragment thereof is extracted by lysing said Vero cells. 
     
     
         6 . The method of  claim 1 , wherein said protective molecule comprises an RNA particle, and said RNA particle comprises said NOI or a fragment thereof. 
     
     
         7 . The method of  claim 1 , wherein said protective molecule comprises an RNA particle, and said RNA particle produces a dsRNA molecule. 
     
     
         8 . The method of  claim 1 , further comprising determining the amount of protective molecule by determining the amount of RNA Particle (RP) titer and providing said protective molecule at a RP titer that when administered protects said animal. 
     
     
         9 . The method of  claim 8  further comprising determining the genome equivalent of the protective molecule, determining the genome equivalent:RNA Particle titer (GE:RP) ratio and providing said protective molecule at a GE:RP ratio that when administered protects said animal. 
     
     
         10 . The method of  claim 1 , wherein said NOI is obtained by a process selected from the group consisting of polymerase chain reaction of said NOI in said sample, and synthesis of said NOI. 
     
     
         11 . The method of  claim 1 , further comprising determine the biotype of said microorganism and producing said vaccine if said microorganism is a biotype different from biotypes of microorganisms for which vaccines exist. 
     
     
         12 . The method of  claim 1 , wherein said at least one animal from which a biological sample is obtained is a first animal in a first plurality of animals exposed to a first microorganism for which a first protective molecule is produced, which first plurality of animals has or will have sufficient exposure to at least one animal or biological sample of a second plurality of animals exposed to a second microorganism, such that said first and second plurality of animals are at risk of exposure to microorganisms present in the other plurality of animals, said method further comprising,
 a) obtaining a biological sample from at least one animal in said second plurality of animals;   b) producing a second protective molecule from a NOI obtained from said biological sample of at least one animal of said second plurality of without the need to isolate said second microorganism; and   c) providing said second protective molecule in said vaccine in an amount that when administered to an animal protects said animal from said biotype of said second microorganism.   
     
     
         13 . The method of  claim 1 , further comprising obtaining biological samples over a period of time, obtaining a NOI from said samples and comparing a later obtained NOI with an earlier obtained NOI and producing said vaccine when said later obtained NOI is different from said earlier obtained NOI. 
     
     
         14 . The method of  claim 1 , wherein said animal is selected from the group consisting of swine, cattle, canines, felines, equines and aquatic invertebrates. 
     
     
         15 . A method of protecting an animal from a biotype of a microorganism, the method comprising,
 a) identifying a nucleic acid molecule of interest or fragment thereof (NOI) of a microorganism, said NOI known to be capable of producing a protective molecule;   b) obtaining a biological sample from at least one animal where said animal has been exposed to said microorganism;   c) obtaining from said sample said NOI of said microorganism without the need to isolate said microorganism;   d) producing a protective molecule from said NOI, said protective molecule, selected from the group consisting of:
 (i) a nucleic acid molecule comprising the sequence of said NOI or a fragment thereof; 
 (ii) a polypeptide or fragment thereof expressed by said NOI; 
 (iii) at least one RNA molecule comprising a sequence that corresponds to all or a portion of said NOI and forms a dsRNA; 
 (iv) at least one RNA molecule that is antisense to all or a portion of said NOI; 
 (v) a nucleic acid molecule producing (iii) or (iv); and 
 (vi) a RNA particle comprising or producing any one of (i)-(v); 
   e) producing a vaccine comprising said protective molecule and wherein said vaccine does not comprise said microorganism or a replicable or living pathogenic microorganism;   f) providing said protective molecule in an amount that when administered to an animal protects said animal from said biotype of said microorganism;   g) administering said protective molecule to at least one animal; and   h) producing a protective response to said biotype of said microorganism in said animal.   
     
     
         16 . The method of  claim 15 , further comprising administering said protective molecule to an animal selected from the group consisting of (i) said animal from which said biological sample was obtained, (ii) an animal living with said animal from which said biological sample was obtained, and (iii) an animal having or which will have sufficient exposure to said animal or a biological sample of (i) or (ii) so as to be at risk of exposure to said biotype of said microorganism. 
     
     
         17 . The method of  claim 15 , further comprising introducing a alphavirus RNA vector comprising said NOI into a host cell, expressing a polypeptide or fragment thereof from said NOI, and extracting said polypeptide or fragment from said host, wherein said extracted polypeptide or fragment thereof comprises said protective molecule and is administered to said animal. 
     
     
         18 . The method of  claim 17 , wherein said vector comprises a Venezuelan Equine Encephalitis (VEE) vector, said host comprises Vero cells and said polypeptide or fragment thereof is extracted by lysing said Vero cells. 
     
     
         19 . The method of  claim 15 , wherein said protective molecule comprises an RNA particle, and said RNA particle comprises said NOI or a fragment thereof. 
     
     
         20 . The method of  claim 15 , wherein said protective molecule comprises an RNA particle, and said RNA particle produces a dsRNA molecule. 
     
     
         21 . The method of  claim 15 , further comprising determining the amount of protective molecule by determining the amount of RNA Particle (RP) titer and providing said protective molecule at a RP titer that when administered protects said animal. 
     
     
         22 . The method of  claim 21  further comprising determining the genome equivalent of the protective molecule, determining the genome equivalent:RNA Particle titer (GE:RP) ratio and providing said protective molecule at a GE:RP ratio that when administered in a vaccine protects said animal. 
     
     
         23 . The method of  claim 15 , wherein said NOI is obtained by a process selected from the group consisting of polymerase chain reaction of said NOI in said sample, and synthesis of said NOI. 
     
     
         24 . The method of  claim 15 , further comprising, determine the biotype of said microorganism and producing and administering said protective molecule if said microorganism is a biotype different from biotypes of microorganisms for which vaccines exist. 
     
     
         25 . The method of  claim 15 , wherein said protective molecule is administered to a plurality of animals living with said animal from which said biological sample was obtained. 
     
     
         26 . The method of  claim 15 , wherein said protective molecule is administered to a single animal, and wherein said single animal is the animal from which said biological sample was obtained. 
     
     
         27 . The method of  claim 15 , wherein said animal from which said biological sample is obtained is an animal in a first plurality of animals exposed to a first microorganism for which a first protective molecule is produced, which first plurality of animals has or will have sufficient exposure to at least one animal or biological sample of a second plurality of animals exposed to a second microorganism, such that said first and second plurality of animals are at risk of exposure to microorganisms present in the other plurality of animals, said method further comprising,
 a) obtaining a biological sample from at least one animal in said second plurality of animals;   b) producing a second protective molecule from a NOI obtained from said biological sample of said at least one animal of said second plurality of without the need to isolate said second microorganism;   c) providing said second protective molecule in an amount that when administered to an animal protects said animal from the biotype of said second microorganism; and   d) administering said first and second protective molecules to said first and second plurality of animals.   
     
     
         28 . The method of  claim 15 , further comprising obtaining biological samples over a period of time, obtaining a NOI from said samples and comparing a later obtained NOI with an earlier obtained NOI and producing said vaccine when said later obtained NOI is different from said earlier obtained NOI. 
     
     
         29 . The method of  claim 15 , wherein said animal is selected from the group consisting of swine, cattle, canines, felines, equines and aquatic invertebrates. 
     
     
         30 . A vaccine to protect an animal from a biotype of a microorganism, said vaccine comprising,
 a) a protective molecule selected from the group consisting of:
 (i) a nucleic acid molecule comprising the sequence of a nucleic acid molecule of interest or a fragment thereof (NOI) known to be capable of producing a protective molecule, said NOI obtained from a sample from an animal which has been exposed to said microorganism; 
 (ii) a polypeptide or fragment thereof expressed by said NOI; 
 (iii) at least one RNA molecule comprising a sequence that corresponds to all or a portion of said NOI and forms a dsRNA; 
 (iv) at least one RNA molecule that is antisense to all or a portion of said NOI; 
 (v) a nucleic acid molecule producing (iii) or (iv); and 
 (vi) a RNA particle comprising or producing any one of (i)-(v); 
   b) said vaccine does not comprise said microorganism or a replicable or living pathogenic microorganism   c) wherein said microorganism is a biotype for which available vaccines when administered to an animal provide no response or a reduced protective response compared to the protective response obtained when administering said protective molecule; and   d) said protective molecule is provided in an amount that when administered protects said animal from said biotype of said microorganism.