Patent
Disclosed is the use of isolated Ats-1 protein in Anaplasma phagocytophilum in the ELISA detection of Anaplasma pathogen. The recombinant expression of Ats-1 and its use as a kit for ELISA are also disclosed.
1. An ELISA kit for the diagnosis of Anaplasma phagocytophilum infection in a human patient, comprising:
a) an isolated recombinant Anaplasma translocated substrate-1 (Ats-1) protein antigen, said isolated recombinant Ats-1 protein antigen having the amino acid sequence of SEQ ID NO: 2, wherein said isolated recombinant Ats-1 protein antigen is immobilized onto the surface of a support made of a material selected from the group consisting of polyethylene, polypropylene and glass;
b) a detection antibody, said detection antibody is an anti-human IgG or anti-human IgM antibody, and said detection antibody is conjugated with a label selected from the group consisting of a fluorophore, radioactive label and enzyme,
wherein
said immobilized isolated recombinant Ats-1 protein antigen binds to an anti-Ats-1 antibody that is present in a biological sample of a human patient suspected of having an infection of Anaplasma phagocytophilium to form a complex of antigen-antibody and detection of the formation of said antigen-antibody complex by said detection antibody is indicative of Anaplasma phagocytophilium infection in said human patient.
2. The ELISA kit of claim 1 , wherein said anti-Ats-1 antibody is an IgG antibody.
3. The ELISA kit of claim 1 , wherein said anti-Ats-1 antibody is an IgM antibody.
4. The ELISA kit of claim 2 , wherein said ELISA kit has a sensitivity of at least 90% and specification of at least 80%.
5. The ELISA kit of claim 3 , wherein said ELISA kit has a sensitivity of at least 90% and specificity of at least 75%.
6. The ELISA kit of claim 1 , wherein said support is made of polypropylene.
7. The ELISA kit of claim 1 , wherein said support is a microtiter well.
8. The ELISA kit of claim 1 , further comprising:
(c) a detection reagent, said detection reagent is an enzyme substrate.
9. The ELISA kit of claim 1 , wherein said fluorophore is fluorescein or rhodamine.
10. The ELISA kit of claim 8 , further comprising:
(d) an instruction, said instruction providing a procedure for detecting the presence of said anti-Ats-1 antibody in a biological sample obtained from a human patient.
11. The ELISA kit of claim 1 , wherein said radioactive label is selected from the group consisting of radioisotope 32 P, 14 C, 125 I, 3 H and 131 I.
12. The ELISA kit of claim 1 , wherein said enzyme is selected from the group consisting of horseradish peroxidase, alkaline phosphatase and glucose oxidase.