Method and composition for determining specific antibody responses to species of filovirus

Abstract

The disclosure relates to compositions, assays, methods and kits comprising one or more amino acid sequences of a filovirus protein, or a fragment thereof, which find use in the detection of a filovirus infection and/or the presence of antibodies specific for a filovirus in a biological sample.

1 . A detection agent comprising one or more amino acid sequences of a filovirus protein, or a fragment thereof, and a substrate, wherein the one or more amino acid sequences of the filovirus protein is attached to the substrate. 
     
     
         2 . The detection agent of  claim 1 , wherein the one or more amino acid sequences of a filovirus protein is from a filovirus selected from  Marburg marburgvirus, Sudan ebolavirus, Zaire ebolavirus, Reston ebolavirus, Bundibugyo ebolavirus , and  Taï Forest ebolavirus.    
     
     
         3 . The detection agent of  claim 1 , wherein the one or more amino acid sequences of a filovirus protein, or fragment thereof, is selected from nucleoprotein (NP), virion protein 40 (VP40), glycoprotein (GP), virion protein (VP35), virion protein (VP30), virion protein (VP24), RNA-dependent RNA polymerase (L), mucin-like domain fragment of GP (GP mucin), GP ectodomain (GPΔTM), or any combination thereof. 
     
     
         4 . The detection agent of  claim 1 , comprising at least three different amino acid sequences of at least three different filovirus proteins, or fragments thereof. 
     
     
         5 . The detection agent of  claim 3 , wherein the one more sequences of a filovirus protein is selected from NP, VP40, and GP, wherein
 NP is selected from a protein having at least 90% sequence identity to the sequence selected from the group consisting of SEQ ID NO:4 (Zaire NP): SEQ ID NO:10 (Sudan NP); SEQ ID NO: 16 (Bundibugyo NP); SEQ ID NO: 22 (Taï Forest NP); SEQ ID: 28 (Reston NP); and SEQ ID NO: 34 (Marburg NP);   VP40 is selected from a protein having at least 90% sequence identity to the sequence selected from the group consisting of SEQ ID NO: 2 (Zaire VP40); SEQ ID NO: 8 (Sudan VP40); SEQ ID NO: 14 (Bundibugyo VP40): SEQ ID 20 (Taï Forest VP40): SEQ ID NO: 26 (Reston VP40); and SEQ ID NO: 32 (Marburg VP40); and   GP is selected from a GP-mucin domain having at least 90% sequence identity to the sequence selected from the group consisting SEQ ID NO: 6 (Zaire GP-mucin); SEQ ID NO: 12 (Sudan GP-mucin); SEQ ID NO: 18 (Bundibugyo GP-mucin): SEQ ID NO: 24 (Taï Forest GP-mucin); SEQ ID NO: 30 (Reston GP-mucin); and SEQ ID NO: 36 (Marburg GP-mucin).   
     
     
         6 . The detection agent of  claim 3 , wherein the one more sequences of a filovirus protein is selected from NP, VP40, and GP, wherein
 NP is selected from the group consisting of SEQ ID NO:4 (Zaire NP); SEQ ID NO:10 (Sudan NP): SEQ ID NO: 16 (Bundibugyo NP); SEQ ID NO: 22 (Taï Forest NP); SEQ ID: 28 (Reston NP); and SEQ ID NO: 34 (Marburg NP);   VP40 is selected from the list consisting of SEQ ID NO: 2 (Zaire VP40); SEQ ID NO: 8 (Sudan VP40); SEQ ID NO: 14 (Bundibugyo VP40); SEQ ID 20 (Taï Forest VP40); SEQ ID NO: 26 (Reston VP40); SEQ ID NO: 32 (Marburg VP40); and   GP comprises a GP-mucin domain selected from the group consisting of SEQ ID NO: 6 (Zaire GP-mucin); SEQ ID NO: 12 (Sudan GP-mucin); SEQ ID NO: 18 (Bundibugyo GP-mucin): SEQ ID NO: 24 (Taï Forest GP-mucin); SEQ ID NO: 30 (Reston GP-mucin); and SEQ ID NO: 36 (Marburg GP-mucin).   
     
     
         7 . The detection agent of  claim 1 , wherein the substrate is selected from the group consisting of a microarray, microparticles, and nanoparticles. 
     
     
         8 . The detection agent of  claim 1 , wherein the substrate is a microarray. 
     
     
         9 . The detection agent of  claim 1 , wherein the one or more amino acid sequences of a filovirus protein is provided as a recombinant protein or a fragment thereof. 
     
     
         10 . A method for detecting the presence of filovirus-specific antibody in biological sample obtained from a subject comprising:
 (a) incubating the biological sample with the detection agent of  claim 1  under conditions that allow binding of the filovirus-specific antibody to the detection agent; and   (b) detecting the filovirus-specific antibody bound to detection agent.   
     
     
         11 . A method for identifying a subject infected with a filovirus comprising determining whether a filovirus-specific antibody is present in a sample obtained from the subject, wherein the determining comprises:
 (a) incubating the biological sample under conditions that allow binding of the filovirus-specific antibody to the detection agent; and   (b) detecting the filovirus-specific antibody bound to the detection agent, wherein the detection of the filovirus-specific antibody identifies that the subject is infected with a filovirus.   
     
     
         12 . A method for making the detection agent of  claim 1  comprising:
 expressing one or more recombinant polynucleotide sequences encoding an amino acid sequence of a filovirus protein, or a fragment thereof in an expression system; and 
 fixing the encoded amino sequence of a filovirus protein, or a fragment thereof, on a surface of the substrate. 
 
     
     
         13 . The method of  claim 12 , wherein the expression system comprises a prokaryotic cell, a eukaryotic cell, or in vitro translation, or any combination thereof. 
     
     
         14 . The method of  claim 13 , wherein the prokaryotic cell comprises  E. coli.    
     
     
         15 . The method of  claim 13 , wherein the eukaryotic cell is selected from the group consisting of yeast, an insect cell, and a mammalian cell.