Diagnosis of
The disclosure, in some aspects, provides a composition comprising labelled and/or tagged and/or bound amino acid sequences useful for the detection of Babesia species. Also disclosed are methods for the detection of infection by one or more Babesia species.
1 . A composition comprising labelled and/or tagged and/or bound amino acid sequences 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, SEQ ID NO: 10, 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, and SEQ ID NO: 19.
2 . The composition of claim 1 , wherein the bound amino acid sequences are bound to a solid support selected from the group consisting of nitrocellulose, nylon, polyvinylidene difluoride (PVDF), plastic, metal, magnetic beads, and agarose.
3 . A method for detecting infection by one or more Babesia species, if present in a biological sample obtained from a subject suspected of having a Babesia infection, the method comprising:
(a) providing the composition of claim 1 ;
(b) providing the biological sample obtained from the subject suspected of having a Babesia infection;
(c) contacting the biological sample with the composition under conditions appropriate for specific antibody binding to an epitope; and
(d) detecting specific binding of IgM- and/or IgG-class antibodies, if present in the biological sample, with the amino acid sequences of the composition, wherein the biological sample is scored as positive for infection by one or more Babesia species when:
(i) a positive immunobinding reaction with IgM-class antibodies is detected for at least two of the amino acid sequences of the composition, or
(ii) a positive immunobinding reaction with IgG-class antibodies is detected for at least two of the amino acid sequences of the composition,
and wherein a positive score for infection indicates infection by one or more Babesia species in the subject.
4 . The method of claim 3 wherein the binding of IgM-class antibodies is detected through the use of an anti-human IgM antibody linked to a detectable moiety.
5 . The method of claim 3 wherein the binding of IgG-class antibodies is detected through the use of an anti-human IgG antibody linked to a detectable moiety.
6 . The method of claim 5 , wherein the detectable moiety is selected from the group consisting of chromophores, radioactive moieties, and enzymes.
7 . The method of claim 6 , wherein the detectable moiety comprises alkaline phosphatase.
8 . The method of claim 6 , wherein the detectable moiety comprises biotin.
9 . The method of claim 3 , wherein the one or more Babesia species comprise one or more of B. microti, B. duncani , B. MO1 , B. divergens, B. venatorum , and B. crassa.
10 . A method for detecting species-specific infection by B. microti and/or B. duncani , if present in a biological sample obtained from a subject suspected of having a Babesia infection, the method comprising:
(a) providing the composition of claim 1 ;
(b) providing the biological sample obtained from the subject suspected of having a Babesia infection;
(c) contacting the biological sample with the composition under conditions appropriate for specific antibody binding to an epitope; and
(d) detecting specific binding of IgM- and/or IgG-class antibodies, if present in the biological sample, with the amino acid sequences of the composition, wherein:
(i) the biological sample is scored as positive for infection by B. microti when a positive immunobinding reaction with IgM- or IgG-class antibodies is detected for at least one of SEQ ID NOs: 1-7, 9, 11, and 13 and at least one of SEQ ID NOs: 8, 10, and 12, or
(ii) the biological sample is scored as positive for infection by B. duncani when a positive immunobinding reaction with IgM- or IgG-class antibodies is detected for at least one of SEQ ID NOs: 1-7, 9, 11, 13, and 16-19 and at least one of SEQ ID NOs: 14 and 15,
and wherein a positive score indicates infection by B. microti and/or B. duncani.
11 . The method of claim 10 wherein the binding of IgM-class antibodies is detected through the use of an anti-human IgM antibody linked to a detectable moiety.
12 . The method of claim 10 wherein the binding of IgG-class antibodies is detected through the use of an anti-human IgG antibody linked to a detectable moiety.
13 . The method of claim 12 , wherein the detectable moiety is selected from the group consisting of chromophores, radioactive moieties, and enzymes.
14 . The method of claim 13 , wherein the detectable moiety comprises alkaline phosphatase.
15 . The method of claim 13 , wherein the detectable moiety comprises biotin.
16 . A vector comprising nucleic acid sequences coding for amino acid sequences 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, SEQ ID NO: 10, 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, and SEQ ID NO: 19.