Peptides, devices, and methods for the detection of anaplasma antibodies

A method for detecting in a sample an antibody to an epitope of an Anaplasma antigen, the method comprising:

• (i) contacting a sample with a composition comprising a population of isolated peptides, said population comprising three or more different peptides, wherein each peptide in the population comprises a sequence selected from:
• (a) SEQ ID NO:
• 1 or a fragment thereof wherein X9 is an amino acid selected from the group consisting of I, P and H, X17 is an amino acid selected from the group consisting of I, W, and Y, X21 is an amino acid selected from the group consisting of R, D, and N, X28 is an amino acid selected from the group consisting of E and N, and X31 is an amino acid selected from the group consisting of L and V;
• (b) SEQ ID NO:
• 2 or a fragment thereof, wherein X3 is an amino acid selected from the group consisting of L, V and A, X7 is an amino acid selected from the group consisting of K, N and Q, X11 is an amino acid selected from the group consisting of R, D, and N, and X15 is an amino acid selected from the group consisting of E, N and Q;
• (c) SEQ ID NO:
• 3 or a fragment thereof wherein X5 is an amino acid selected from the group consisting of V and A, X7 is an amino acid selected from the group consisting of G, I and H, X11 is an amino acid selected from the group consisting of E, N, and Q, X18 is an amino acid selected from the group consisting of D and N, X21 is an amino acid selected from the group consisting of R, D, and N, X25 is an amino acid selected from the group of Q, D, and E, X28 is an amino acid selected from the group consisting of and E or N, X31 is an amino acid selected from the group consisting of L and V, X45 is an amino acid selected from the group consisting of K and Q, X48 is an amino acid selected from the group consisting of F and V, X51 is an amino acid selected from the group consisting of D and N, X54 is an amino acid selected from the group consisting of E and Q, X57 is an amino acid selected from the group consisting of S and Q, X60 is an amino acid selected from the group consisting of F and W, X63 is an amino acid selected from the group consisting of I and V, and X66 is an amino acid selected from the group consisting of Q and D;
• (d) SEQ ID NO:
• 4 or a fragment thereof, wherein X6 is an amino acid selected from the group consisting of K and Q, X9 is an amino acid selected from the group consisting of F and V, X12 is an amino acid selected from the group consisting of D and N, X15 is an amino acid selected from the group consisting of E and Q, X18 is an amino acid selected from the group consisting of S and Q, X21 is an amino acid selected from the group consisting of F and W, X24 is an amino acid selected from the group consisting of I and V, and X27 is an amino acid selected from the group consisting of Q and D;
• (e) SEQ ID NO:
• 5 or a fragment thereof, wherein X2 is an amino acid selected from the group consisting of I and V, X10 is an amino acid selected from the group consisting of S and Y, and X23 is an amino acid selected from the group consisting of E and N;
• (f) SEQ ID NO:
• 6 or a fragment thereof;
• (g) SEQ ID NO:
• 7 or a fragment thereof, wherein X5 is an amino acid selected from the group consisting of S and Q, X9 is an amino acid selected from the group consisting of F and Y, X13 is an amino acid selected from the group consisting of R and H, X16 is an amino acid selected from the group consisting of W and Y, X18 is an amino acid selected from the group consisting of S and Q, X22 is an amino acid selected from the group consisting of K and H, and X27 is an amino acid selected from the group consisting of N and D;
• (h) SEQ ID NO:
• 8 or a fragment thereof, wherein X10 is an amino acid selected from the group consisting of V, L and I, and X11 is an amino acid selected from the group consisting of A and L;
• and
• (i) SEQ ID NO:
• 9 or a fragment thereof, wherein X5 is an amino acid selected from the group consisting of V and A, X7 is an amino acid selected from the group consisting of G, I and H, X11 is an amino acid selected from the group consisting of E, N, and Q, X21 is an amino acid selected from the group consisting of R, D, and N, X25 is an amino acid selected from the group consisting of Q, D, and E, X28 is an amino acid selected from the group consisting of E and N, and X31 is an amino acid selected from the group consisting of L and V;
• and
• (ii) detecting formation of an antibody-peptide complex comprising one or more peptides in the composition, wherein formation of said complex is indicative of an antibody to an epitope of an Anaplasma antigen being present in said sample.

A method for diagnosing anaplasmosis in a subject, the method comprising:

• (i) contacting a sample from the subject with a composition comprising a population of isolated peptides, said population comprising three or more different peptides, wherein each peptide in the population comprises a sequence selected from:
• (a) SEQ ID NO:
• 1 or a fragment thereof, wherein X9 is an amino acid selected from the group consisting of I, P and H, X17 is an amino acid selected from the group consisting of I, W, and Y, X21 is an amino acid selected from the group consisting of R, D, and N, X 28 is an amino acid selected from the group consisting of E and N, and X31 is an amino acid selected from the group consisting of L and V;
• (b) SEQ ID NO:
• 2 or a fragment thereof, wherein X3 is an amino acid selected from the group consisting of L, V and A, X7 is an amino acid selected from the group consisting of K, N and Q, X11 is an amino acid selected from the group consisting of R, D, and N, and X15 is an amino acid selected from the group consisting of E, N and Q;
• (c) SEQ ID NO:
• 3 or a fragment thereof, wherein X5 is an amino acid selected from the group consisting of V and A, X7 is an amino acid selected from the group consisting of G, I and H, X11 is an amino acid selected from the group consisting of E, N, and Q, X18 is an amino acid selected from the group consisting of D and N, X21 is an amino acid selected from the group consisting of R, D, and N, X25 is an amino acid selected from the group consisting of Q, D, and E, X28 is an amino acid selected from the group consisting of and E or N, X31 is an amino acid selected from the group consisting of L and V, X45 is an amino acid selected from the group consisting of K and Q, X48 is an amino acid selected from the group consisting of F and V, X51 is an amino acid selected from the group consisting of D and N, X54 is an amino acid selected from the group consisting of E and Q, X57 is an amino acid selected from the group consisting of S and Q, X60 is an amino acid selected from the group consisting of F and W, X63 is an amino acid selected from the group consisting of I and V, and X66 is an amino acid selected from the group consisting of Q and D;
• (d) SEQ ID NO:
• 4 or a fragment thereof, wherein X6 is an amino acid selected from the group consisting of K and Q, X9 is an amino acid selected from the group consisting of F and V, X12 is an amino acid selected from the group consisting of D and N, X15 is an amino acid selected from the group consisting of E and Q, X18 is an amino acid selected from the group consisting of S and Q, X21 is an amino acid selected from the group consisting of F and W, X24 is an amino acid selected from the group consisting of I and V, and X27 is an amino acid selected from the group consisting of Q and D;
• (e) SEQ ID NO:
• 5 or a fragment thereof, wherein X2 is an amino acid selected from the group consisting of I and V, X10 is an amino acid selected from the group consisting of S and Y, and X23 is an amino acid selected from the group consisting of E and N;
• (f) SEQ ID NO:
• 6 or a fragment thereof;
• (g) SEQ ID NO:
• 7 or a fragment thereof wherein X5 is an amino acid selected from the group consisting of S and Q, X9 is an amino acid selected from the group consisting of F and Y, X13 is an amino acid selected from the group consisting of R and H, X16 is an amino acid selected from the group consisting of W and Y, X18 is an amino acid selected from the group consisting of S and Q, X22 is amino acid selected from the group consisting of K and H, and X27 is an amino acid selected from the group consisting of N and D;
• (h) SEQ ID NO:
• 8 or a fragment thereof, wherein X10 is an amino acid selected from the group consisting of V, L and I, and X11 is an amino acid selected from the group consisting of A and L;
• and
• (i) SEQ ID NO:
• 9 or a fragment thereof, wherein X5 is an amino acid selected from the group consisting of V and A, X7 is an amino acid selected from the group consisting of G, I and H, X11 is an amino acid selected from the group consisting of E, N, and Q, X21 is an amino acid selected from the group consisting of R, D, and N, X25 is an amino acid selected from the group consisting of Q, D, and E, X28 is an amino acid selected from the group consisting of E and N, and X31 is an amino acid selected from the group consisting of L and V;
• and
• (ii) detecting formation of an antibody-peptide complex comprising one or more peptides in the composition,
• wherein formation of the complex is indicative of the subject having anaplasmosis.

A method for identifying the species of Anaplasma infecting a subject, the method comprising:

• (a) contacting a sample from the subject with a first population of isolated peptides and a second population of isolated peptides, wherein the first population of isolated peptides specifically binds to antibodies against antigens from multiple Anaplasma species, wherein the second population of isolated peptides specifically binds to antibodies against antigens from a single Anaplasma species;
• (b) detecting formation of a first antibody-peptide complex comprising one or more peptides in the first population;
• and
• (c) detecting formation of a second antibody-peptide complex comprising one or more peptides in the second population, wherein formation of both the first and second antibody-peptide complexes indicates that the subject is infected with the Anaplasma species that is specifically bound by the second population of isolated peptides.

A method for identifying the species of Anaplasma infecting a subject, the method comprising:

• (a) contacting a sample from the subject with a first population of isolated peptides and a cell extract of a single Anaplasma species, wherein the first population of isolated peptides specifically binds to antibodies against antigens from multiple Anaplasma species;
• (b) detecting formation of a first antibody-peptide complex comprising one or more peptides in the first population;
• and
• (c) detecting formation of an antibody-cell extract complex comprising one or more components in the cell extract, wherein formation of both the first antibody-peptide complex and the antibody-cell extract complex indicates that the subject is infected with the Anaplasma species that produced the cell extract.