Type IV secretion system proteins in sero-detection of
Disclosed are two (2) proteins in the Type IV Secretion System (TIVSS) in Anaplasma phagocytophilum (namely, virB10 and virB11) useful in the ELISA detection of Anaplasma pathogen. The recombinant expression of virB10 and virB11 and their use as kits for ELISA are also disclosed.
1. A method of detecting the presence of an antibody against Anaplasma phagocytophilum in a biological sample of a mammal, comprising the steps of:
(i) immobilizing an isolated polypeptide onto a surface, wherein said isolated polypeptide consists of SEQ ID NO: 32 or SEQ ID NO: 33;
(ii) contacting said isolated polypeptide with a patient's biological sample, under conditions that allow formation of an antibody-antigen complex, wherein said biological sample is suspected of containing an antibody against Anaplasma phagocytophilum ; and
(iii) detecting the formation of said antibody-antigen complex, wherein detection of said antibody-antigen complex is indicative of the presence of said antibody against Anaplasma phagocytophilum in said biological sample.
2. The method of claim 1 , wherein said mammal is a human.
3. The method of claim 1 , wherein said antibody is an IgG or IgM.
4. The method of claim 1 , wherein said method is an enzyme-linked immunosorbent assay (ELISA).
5. The method of claim 4 , wherein said enzyme-linked immunosorbent assay (ELISA) has a sensitivity of at least >70%.
6. The method of claim 4 , wherein said enzyme-linked immunosorbent assay (ELISA) has a specificity of at least 80%.
7. A method of diagnosing an infection of Anaplasma phagocytophilum in a mammal, comprising the steps of:
(i) obtaining a biological sample from a mammal suspected of having a Anaplasma phagocytophilum infection;
(ii) immobilizing an isolated polypeptide on to a surface, wherein said isolated polypeptide consists of SEQ ID NO: 32 or SEQ ID NO: 33;
(iii) contacting said isolated polypeptide with said biological sample, under conditions that allow formation of an antibody-antigen complex; and
(iv) detecting said antibody-antigen complex, wherein said detected antibody-antigen complex is indicative of the presence of said antibody against Anaplasma phagocytophilum in said biological sample.
8. The method of claim 7 , wherein said isolated polypeptide consists of SEQ ID NO: 32.
9. The method of claim 7 , wherein said isolated polypeptide consists of SEQ ID NO: 33.
10. The method of claim 7 , wherein said mammal is a human.
11. The method of claim 7 , wherein said biological sample is whole blood.
12. The method of claim 7 , wherein said antibody is IgG or IgM.
13. The method of claim 7 , wherein said contacting step is performed at room temperature for about 1 hour.