Treponema pallidum triplet antigen
View Patent ↗A Treponema pallidum triplet antigen construct is disclosed which includes three Treponema pallidum antigens (TP15, TP17, and TP47), as well as a ten amino acid leader sequence (tag 261) and human copper zinc superoxide dismutase (hSOD). This construct is optimized for in vitro diagnosis of syphilis infection. Plasmids containing DNA encoding the triplet antigen, host cells, production methods, detection methods, and kits are also disclosed.
1. A Treponema pallidum triplet antigen comprising the amino acid sequence of SEQ ID NO:11.
2. The Treponema pallidum triplet antigen of claim 1 labeled with a detectable marker.
3. The Treponema pallidum triplet antigen of claim 2 wherein the detectable marker is horseradish peroxidase.
4. A method of detecting the presence of Treponema pallidum antibodies in a sample, the method comprising:
contacting the sample with a first antigen which is immobilized on a solid support and which comprises a Treponema pallidum triplet antigen comprising SEQ ID NO: 11, wherein Treponema pallidum antibodies present in the sample bind to the Treponema pallidum triplet antigen forming an antibody/antigen complex; and
detecting the antibody/antigen complex, thereby detecting the presence of Treponema pallidum antibodies, wherein the antibody/antigen complex is detected by
(i) contacting the antibody/antigen complex with a Treponema pallidum triplet antigen which comprises SEQ ID NO: 1, is labeled with a detectable marker and binds to the antibody present in the antibody/antigen complex forming an antigen/antibody/labeled antigen complex, and detecting the antigen/antibody/labeled antigen complex; or
(ii) contacting the antibody/antigen complex with a labeled anti-human antibody, wherein the labeled anti-human antibody binds to the antibody present in the antibody/antigen complex forming an antigen/antibody/labeled anti-antibody complex; and detecting the antigen/antibody/labeled anti-antibody complex.
5. A kit for detection of Treponema pallidum antibodies, the kit comprising:
a first antigen comprising the Treponema pallidum triplet antigen of claim 1 ; and
a second antigen comprising the Treponema pallidum triplet antigen of claim 1 , wherein the second antigen is labeled with a detectable marker.
6. The kit of claim 5 further comprising positive or negative controls.
7. The kit of claim 5 further comprising calibration samples.
8. The kit of claim 5 further comprising one or more of enzyme conjugate, substrate for enzyme conjugate, buffer solution, and washing solution.