Detection kit containing a novel recombinant 15-kDA polypeptide useful for detecting human infection with
Disclosed are the cloning and expression of a novel antigen of Bartonella henselae . The recombinant polypeptide is found to be highly immunogenic and is useful as a diagnostic test antigen. The polypeptide of the present invention provides the basis of a diagnostic assay that is sensitive, rapid and accurate diagnosis of infection with Bartonella henselae using patient's sera. Disclosed also are the ELISA for both IgG and IgM and allows diagnosis of early and late infection.
1. A composition for detection of the presence of anti-15-kDa IgG or IgM antibodies in a human suspected of Bartonella henselae infection, comprising:
a) an isolated recombinant 15-kDa polypeptide, said isolated recombinant 15-kDa polypeptide having the amino acid sequence of SEQ ID NO: 2; and
b) a support,
wherein said isolated recombinant 15-kDa polypeptide is immobilized on said support and is prepared by a process, said process comprises the steps of:
i) introducing a 15-kDa insert DNA having a nucleotide sequence of SEQ ID NO. 1 in an E. coli cell;
ii) growing said E. coli cell that has said 15-kDa insert DNA in a culture medium;
iii) adding 5-10 mg/mL glucose followed by IPTG in said culture medium; and
iv) isolating said recombinant 15-kDa polypeptide from said culture medium.
2. The composition of claim 1 , wherein said support is selected from the group consisting of polyethylene, polypropylene and glass.
3. The composition of claim 2 , wherein said support is polypropylene.
4. The composition of claim 1 , wherein said support is a microtiter well.
5. The composition of claim 1 , further comprising:
c) packaging material.
6. The composition of claim 5 , wherein said packaging material comprises an instruction for detecting the presence of an antibody against Bartonella henselae.
7. The composition of claim 1 , wherein said 15-kDa insert DNA in step i) is contained in a vector.
8. The composition of claim 7 , wherein said vector comprises a promoter of DNA transcription operably linked to said 15-kDa insert DNA.
9. The composition of claim 8 , wherein said vector is selected from the group consisting of pET, pENTR and pCR 8/GW/TOPO and said promoter is selected from the group consisting of lac promoter, trp promoter and tac promoter.
10. The composition of claim 9 , wherein said vector is pET and said promoter is lac promoter.
11. The composition of claim 1 , wherein said E. coli cell is NovaBlue K12 strain, BL21 (DE3) or BL21 pLyss (DE3).