Patent
Disclosed are the cloning and expression of novel antigens in Babesia microti . The recombinant polypeptides are highly immunogenic. The polypeptides of the present invention provide the basis of a diagnostic assay that is sensitive, rapid and accurate using patient's sera. Also disclosed is an IgG and IgM ELISA using two novel recombinant antigens in the diagnosis of Babesia infection.
1. A method of detecting the presence of an antibody against Babesia microti in a biological sample of a mammal, comprising:
(i) immobilizing an isolated polypeptide set forth as SEQ ID NO: 2 or SEQ ID NO. 4 onto a surface;
(ii) contacting said isolated polypeptide with a patient's biological sample, under conditions that allow formation of an antibody-antigen complex, said biological sample containing either IgG or IgM;
(iii) detecting the formation of said antibody-antigen complex, wherein said detected antibody-antigen complex is indicative of the presence of said antibody against Babesia microti in said biological sample.
2. The method of claim 1 , wherein the 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 the detecting step further comprises adding an indicator reagent comprising a signal generating compound after step (ii).
5. The method of claim 4 , wherein said indicator reagent is horseradish peroxidase.
6. A method of diagnosing an infection of Babesia microti in a mammal, comprising:
(i) obtaining a biological sample from a mammal suspected of having a Babesia microti infection;
(ii) immobilizing an isolated polypeptide set forth as SEQ ID NO: 2 or SEQ ID NO: 4 onto a surface;
(iii) contacting said purified polypeptide with said biological sample, under conditions that allow formation of antibody-antigen complex;
(iv) detecting said antibody-antigen complex, wherein said detected antibody-antigen complex is indicative of the presence of said antibody against Babesia microti in said biological sample.
7. The method of claim 6 , wherein the mammal is a human.
8. The method of claim 6 wherein said antibody is selected from an isotope consisting of IgG and IgM.
9. The method of claim 6 , wherein the contacting step is performed at room temperature for about 1 hour.
10. The method of claim 9 , wherein the detecting step further comprises adding an indicator reagent comprising a signal generating compound after step (iii).
11. The method of claim 10 , wherein said indicator reagent is horseradish peroxidase.