Extraction reagent for use in an assay for detection of group A
An enzymatic extraction agent, as well as methods, compositions and kits for detecting Group A Streptococcus in a biological sample, which involve the enzymatic agent, are described.
1. An enzymatic extraction agent comprising a recombinant multimeric PlyC holoenzyme consisting of PlyCA and PlyCB gene products and a labeled antibody that specifically binds to a Strep A antigen.
2. The enzymatic extraction agent of claim 1 , wherein the antibody is coupled to microbeads, gold sol, or carbon microparticles.
3. A device for detecting the presence of Group A Streptococcus in a test sample, the device comprising:
a matrix having (i) a sample receiving zone for receiving a test sample containing or suspected of containing a Group A Streptococcus -specific antigen, wherein the test sample comprises is the biological sample and an enzymatic extraction agent comprising a recombinant multimeric PlyC holoenzyme consisting of PlyCA and PlyCB gene products and a labeled antibody that specifically binds to the Group A Streptococcus -specific antigen and (ii) a capture zone having means for specifically binding a labeled Group A Streptococcus -specific antigen thereon, wherein the sample receiving zone and the capture zone are arranged on the matrix in a liquid flow path.
4. The device of claim 3 , wherein the sample receiving zone comprises N-acetyl-D-glucosamine (NAG).
5. The device of claim 3 , wherein the antibody is a polyclonal, monoclonal, or recombinant chimeric antibody coupled to fluorescent microbeads.
6. The device of claim 3 , wherein the antibody is coupled to europium chelate-impregnated microbeads.
7. The device of claim 3 , wherein the means for specifically binding the labeled Group A Streptococcus -specific antigen is a capture antibody.
8. The device of claim 7 , wherein the capture antibody is a polyclonal, monoclonal, or recombinant chimeric antibody.
9. A kit comprising:
a device comprising a matrix having (i) a sample receiving zone for receiving a test sample containing or suspected of containing a Group A Streptococcus -specific antigen and (ii) a capture zone having means for specifically binding a labeled Group A Streptococcus -specific antigen thereon, wherein the sample receiving zone and the capture zone are arranged on the matrix in a liquid flow path; and
a container comprising the enzymatic extraction agent of claim 1 .
10. The kit of claim 9 , wherein the antibody is coupled to europium chelate-impregnated microbeads.
11. The kit of claim 9 , wherein at least one of the enzymatic extraction agent or the sample receiving zone comprise N-acetyl-D-glucosamine (NAG).
12. The kit of claim 11 , wherein the NAG is deposited on the sample receiving zone.
13. A kit comprising:
a device comprising a matrix having (i) a sample receiving zone for receiving a test sample containing or suspected of containing a Group A Streptococcus -specific antigen, wherein the test sample comprises a biological sample and the enzymatic extraction agent of claim 1 and (ii) a capture zone having means for specifically binding a labeled Group A Streptococcus -specific antigen thereon, wherein the sample receiving zone and the capture zone are arranged on the matrix in a liquid flow path;
a container comprising the enzymatic extraction agent of claim 1 ;
an instrument for collecting the biological sample; and
instructions for use.
14. The kit of claim 13 , wherein the instrument is a swab.
15. The kit of claim 13 , wherein the instructions do not caution to not touch tongue, sides or top of mouth with the instrument when collecting a sample.
16. The enzymatic extraction agent of claim 2 , wherein the microbeads are fluorescent microbeads, europium chelate-impregnated microbeads, colored microbeads, or magnetic microbeads.
17. The enzymatic extraction agent of claim 1 , wherein the extraction agent is in the form of a liquid, lyophilized or dried composition.
18. The enzymatic extraction agent of claim 17 , wherein the extraction agent is in the form of a liquid and the recombinant PlyC holoenzyme is present at a concentration from 0.1 μg/mL to 80 μg/mL.
19. The kit of claim 13 , wherein the enzymatic extraction agent is in the form of a liquid and the recombinant PlyC holoenzyme is present at a concentration from 0.1 μg/mL to 80 μg/mL.
20. The enzymatic extraction agent of 1 , wherein the recombinant multimeric PlyC holoenzyme consists of a single PlyCA protein bound to the PlyCB octamer.
21. The device of claim 3 , wherein the biological sample is collected on a swab.
22. The device of claim 3 , wherein the biological sample is a liquid sample.
23. The device of claim 22 , wherein the liquid sample is a body fluid.
24. The device of claim 23 , wherein the body fluid is urine, saliva, sputum, mucus, plasma, serum, amniotic fluid, semen, vaginal secretion, tear, spinal fluid, or body washing.
25. The device of claim 3 , wherein the biological sample is a solid or semi-solid sample.
26. The device of claim 25 , wherein the solid or the semi-solid sample is feces, biopsy specimen, or skin.
27. The device of claim 21 , wherein the sample is eluted from the swab into a transfer medium.