Detection of Lyme disease
The disclosure, in some aspects, provides antigen-specific amino acid sequences for Borrelia burgdorferi sensu lato species.
1. A panel for detecting IgM- or IgG-class antibodies, the panel comprising:
a C6 chimera consisting of SEQ ID NO: 45; and
a plurality of groups of labelled and/or tagged and/or bound amino acid sequences, wherein labelled and/or tagged amino acid sequence is labelled and/or tagged with a detectable moiety, and wherein the plurality of groups of labelled and/or tagged and/or bound amino acid sequences is selected from the following groups:
(i) SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, and SEQ ID NO: 10,
(ii) SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, and SEQ ID NO: 26,
(iii) SEQ ID NO: 32 and SEQ ID NO: 33,
(iv) SEQ ID NO: 27 and SEQ ID NO: 28;
(v) SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, and SEQ ID NO: 20,
(vi) SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, and SEQ ID NO: 39,
(vii) SEQ ID NO: 29,
(viii) SEQ ID NO: 30,
(ix) SEQ ID NO: 31,
(x) SEQ ID NO: 40,
(xi) SEQ ID NOs: 41 and 42, and
(xii) SEQ ID NO: 43.
2. The panel of claim 1 , wherein the labelled and/or tagged and/or bound amino acid sequences are bound to a substance selected from the group consisting of nitrocellulose, nylon, polyvinylidene difluoride (PVDF), magnetic beads, and agarose.
3. The panel of claim 1 further comprising one or more of SEQ ID NO: 44 and SEQ ID NO: 46.
4. The panel of claim 1 , wherein each of the one or more amino acid sequences are tagged with an antibody with specificity for the amino acid sequence.
5. A method for detecting IgM- or IgG-class antibodies resulting from infection by one or more Borrelia burgdorferi sensu lato (Bbsl) species, if present in a biological sample obtained from a subject suspected of having Lyme disease, the method comprising:
(a) providing a screening panel comprising:
a C6 chimera consisting of SEQ ID NO: 45; and
a plurality of groups of labelled and/or tagged and/or bound amino acid sequences, wherein the plurality of groups of labelled and/or tagged and/or bound amino acid sequences is selected from the following groups:
(i) SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, and SEQ ID NO: 10,
(ii) SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, and SEQ ID NO: 26,
(iii) SEQ ID NO: 32 and SEQ ID NO: 33,
(iv) SEQ ID NO: 27 and SEQ ID NO: 28;
(v) SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, and SEQ ID NO: 20,
(vi) SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, and SEQ ID NO: 39,
(vii) SEQ ID NO: 29,
(viii) SEQ ID NO: 30,
(ix) SEQ ID NO: 31,
(x) SEQ ID NO: 40,
(xi) SEQ ID NOs: 41 and 42, and
(xii) SEQ ID NO: 43;
(b) providing the biological sample obtained from the subject suspected of having Lyme disease;
(c) contacting the biological sample with the screening panel of step (a) under conditions appropriate for specific antibody binding to an epitope; and
(d) detecting specific binding of IgM- or IgG-class antibodies, if present in the biological sample, with amino acid sequences included in the selected groups of the screening panel of step (a), wherein the sample is scored as positive for infection by one or more Bbsl species when:
(1) a positive immunobinding reaction with IgM-class antibodies is detected for SEQ ID NO: 45 and for at least one amino acid sequence from at least two of groups (i)-(iii),
(2) a positive immunobinding reaction with IgG-class antibodies is detected for SEQ ID NO: 45 and for at least one amino acid sequence from at least five of groups (i)-(iv) and (vii)-(xii),
(3) a positive immunobinding reaction with IgM-class antibodies is detected for SEQ ID NO: 45 and for at least one amino acid sequence from at least two of groups (i)-(iii) and (v), or
(4) a positive immunobinding reaction with IgG-class antibodies is detected for SEQ ID NO: 45 and for at least one amino acid sequence from at least two of groups (i)-(vi),
and wherein a positive score for infection indicates the presence of antibodies to one or more Bbsl species in the subject.
6. The method of claim 5 wherein the binding of IgM-class antibodies is detected through the use of an anti-human IgM antibody linked to a detectable moiety.
7. The method of claim 5 wherein the binding of IgG-class antibodies is detected through the use of an anti-human IgG antibody linked to a detectable moiety.
8. The method of claim 6 or 7 , wherein the detectable moiety is selected from the group consisting of chromophores, radioactive moieties, and enzymes.
9. The method of claim 8 , wherein the detectable moiety comprises alkaline phosphatase.
10. The method of claim 8 , wherein the detectable moiety comprises biotin.
11. The method of claim 5 , wherein the one or more Bbsl species comprise B. afzelii, B. garinii, B. californiensis, B. spielmanii, B. mayonii, B. valaisiana, B. bavariensis, B. burgdorferi B31, and B. burgdorferi 297.
12. The method of claim 5 , wherein the screening panel of step (a) further comprises at least one or more of SEQ ID NO: 44 and SEQ ID NO: 46.
13. A method for detecting IgM-class and IgG-class antibodies resulting from infection by one or more Borrelia burgdorferi sensu lato (Bbsl) species, if present in a biological sample obtained from a subject suspected of having Lyme disease, the method comprising:
(a) providing a screening panel comprising:
a C6 chimera consisting of SEQ ID NO: 45; and
a plurality of groups of labelled and/or tagged and/or bound amino acid sequences, wherein the plurality of groups of labelled and/or tagged and/or bound amino acid sequences is selected from the following groups:
(i) SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, and SEQ ID NO: 10,
(ii) SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, and SEQ ID NO: 26,
(iii) SEQ ID NO: 32 and SEQ ID NO: 33,
(iv) SEQ ID NO: 27 and SEQ ID NO: 28;
(v) SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, and SEQ ID NO: 20,
(vi) SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, and SEQ ID NO: 39,
(vii) SEQ ID NO: 29,
(viii) SEQ ID NO: 30,
(ix) SEQ ID NO: 31,
(x) SEQ ID NO: 40,
(xi) SEQ ID NOs: 41 and 42, and
(xii) SEQ ID NO: 43;
(b) providing the biological sample obtained from the subject suspected of having Lyme disease;
(c) contacting the biological sample with the screening panel of step (a) under conditions appropriate for specific antibody binding to an epitope; and
(d) detecting specific binding of IgM-class and IgG-class antibodies, if present in the biological sample, with amino acid sequences included in the selected groups of the screening panel of step (a), wherein the sample is scored as positive for infection by one or more Bbsl species when a positive immunobinding reaction with IgM-class or IgG-class antibodies is detected for SEQ ID NO: 45 and:
(1) (A) a positive immunobinding reaction with IgM-class antibodies is detected for at least one amino acid sequence from at least two of groups (i)-(iii), and
(1) (B) a positive immunobinding reaction with IgG-class antibodies is detected for at least one amino acid sequence from at least five of groups (i)-(iv) and (vii)-(xii); or
(2) (A) a positive immunobinding reaction with IgM-class antibodies is detected for at least one amino acid sequence from at least two of groups (i)-(v), and
(2) (B) a positive immunobinding reaction with IgG-class antibodies is detected for at least one amino acid sequence from at least two of groups (i)-(vi),
and wherein a positive score for infection indicates the presence of antibodies to one or more Bbsl species in the subject.
14. The method of claim 13 wherein the binding of IgM-class antibodies is detected through the use of an anti-human IgM antibody linked to a detectable moiety.
15. The method of claim 13 wherein the binding of IgG-class antibodies is detected through the use of an anti-human IgG antibody linked to a detectable moiety.
16. The method of claim 14 or 15 , wherein the detectable moiety is selected from the group consisting of chromophores, radioactive moieties, and enzymes.
17. The method of claim 16 , wherein the detectable moiety comprises alkaline phosphatase.
18. The method of claim 16 , wherein the detectable moiety comprises biotin.
19. The method of claim 13 , wherein the one or more Bbsl species comprise B. afzelii, B. garinii, B. californiensis, B. spielmanii, B. mayonii, B. valaisiana, B. bavariensis, B. burgdorferi B31, and B. burgdorferi 297.
20. The method of claim 13 , wherein the screening panel of step (a) further comprises at least one or more of SEQ ID NO: 44 and SEQ ID NO: 46.