Replies to post #12966 on NanoLogix Inc (NNLX)

[Omar8]

Within Couple more weeks more news oh boy....





"We are pleased to announce the patent for rapid viral testing and diagnosis is scheduled for grant and issuance by the United States Patent and Trademark Office (USPTO) in less than two weeks. Information on the patent should be published in the USPTO Official Gazette for Patents on November 24th. Prior to that date both an international patent application under the Patent Cooperation Treaty (PCT) and a Continuation In Part (CIP) will be filed to broaden both the geographic and technological scope of the Intellectual Property protection and to add additional aspects to the technology that have been discovered or invented since the original application date.


From Oct 14.....I wonder if the news will include what was mention below ? Does grant and issuance mean issued ?




Upon successful results for work currently being performed by a third party to apply NanoLogix technology to a separate testing platform for COVID19 detection, other applications may follow. Once the patent is issued we will reveal the broad and detailed extent of allowed claims for this technology.

[Omar8]

International Patent




Why is international Patent being done so much later than the one in US ?



"Rapid COVID19 test Patent Update

Patent filed May 18 Grant and Issuance will Occur within Two Weeks

We are pleased to announce the patent for rapid viral testing and diagnosis is scheduled for grant and issuance by the United States Patent and Trademark Office (USPTO) in less than two weeks. Information on the patent should be published in the USPTO Official Gazette for Patents on November 24th. Prior to that date both an international patent application under the Patent Cooperation Treaty (PCT) and a Continuation In Part (CIP) will be filed to broaden both the geographic and technological scope of the Intellectual Property protection and to add additional aspects to the technology that have been discovered or invented since the original application date.

Full details of the granted patent, with the complete list of twenty-five granted claims will be available upon Gazette publication.

[soupoftheday]

I FOUND THE PATENT!!! >>>

[soupoftheday]

Twenty-five granted claims >>>

What is claimed is:

1. A method for rapid, highly specific and sensitive, detection and quantification of a virus by observing binding of a viral substrate to its host receptor protein, comprising the steps of: coating a plurality of microtiter wells in a microtiter plate with a host receptor protein contained in a coating buffer; incubating the plurality of microtiter wells overnight; washing the microtiter wells; adding a blocking solution to the plurality of microtiter wells; washing the plurality of microtiter wells three times; adding a viral substrate to the plurality of microtiter wells; incubating the plurality of microtiter wells for 20 minutes; washing the plurality of microtiter wells three times; adding an antibody directed against the viral substrate to the plurality of microtiter wells; incubating the plurality of microtiter wells for 20 minutes; adding a horseradish peroxidase (HRP)-conjugated antibody directed against the antibody to the plurality of microtiter wells; incubating the plurality of microtiter wells for 20 minutes; washing the plurality of microtiter wells three times; adding a TMB solution to the plurality of microtiter wells; adding a stop solution to the plurality of microtiter wells; and detecting the viral substrate in the microtiter wells by observing those microtiter wells that undergo a color change, or quantifying the concentration of the viral substrate by reading optical density at 450 nm, wherein the method is completed by a user in about one hour.

2. The rapid assay of claim 1, wherein the host receptor protein and the viral substrate that binds to its host receptor protein is selected from SARS-CoV-2: Spike protein and angiotensin-converting enzyme 2 (ACE2), SARS-CoV-2: Spike protein and other host protein candidates, Betacoronaviruses (lineage A):Hemagglutinin (HA) esterase and sialic acid receptors, Influenza:HA protein, sialic acid receptors and HA2; Murine hepatitis virus (MHV): Spike protein and carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1), and Middle East respiratory syndrome (MERS): Spike protein and dipeptidyl peptidase 4 (DPP4/CD26).

3. The rapid method of claim 2, wherein the host receptor protein and the viral substrate that binds to its host receptor protein is ACE2 and SARS-CoV-2 Spike protein.

4. The rapid method of claim 1, wherein the antibody is a rabbit polyclonal antibody.

5. The rapid method of claim 1, wherein the HRP-conjugated antibody is an HRP-conjugated anti-rabbit polyclonal goat antibody.

6. The rapid method of claim 1, wherein after adding the blocking solution to the microtiter wells, the microtiter plate can be stored, after which it can be shipped to a user at another site, as the assay start time begins only when the viral substrate is added to the microtiter wells.

7. A method for rapid, highly specific and sensitive, detection and quantification of a virus in an individual suspected of being infected with a virus by observing binding of a specimen taken from the individual with a host receptor protein, comprising the steps of: coating a plurality of microtiter wells with a host receptor protein contained in a coating buffer; incubating the plurality of microtiter wells overnight; washing the microtiter wells; adding a blocking solution to the plurality of microtiter wells; washing the plurality of microtiter wells three times; adding a viral substrate to the plurality of microtiter wells; incubating the plurality of microtiter wells for 20 minutes; washing the plurality of microtiter wells three times; adding an antibody directed against the viral substrate to the plurality of microtiter wells; incubating the plurality of microtiter wells for 20 minutes; adding a horseradish peroxidase (HRP)-conjugated antibody directed against the antibody to the plurality of microtiter wells; incubating the plurality of microtiter wells for 20 minutes; washing the plurality of microtiter wells three times; adding a TMB solution to the plurality of microtiter wells; adding a Stop solution to the plurality of microtiter wells; and detecting the viral substrate in the microtiter wells by observing those microtiter wells that undergo a color change, or quantifying the concentration of the viral substrate by reading optical density at 450 nm, wherein the method is completed by a user in about one hour.

8. The rapid method of claim 7, wherein after adding the blocking solution to the microtiter wells, the microtiter plate can be stored, after which it can be shipped to a user at another site, as the assay start time begins only when the viral substrate is added to the microtiter wells.

9. The rapid method of claim 7, wherein the suspected virus is SARS-CoV-2 and the host receptor protein is ACE2.

10. The rapid method of claim 7, wherein the infection is consistent with COVID-19.

11. The rapid method of claim 7, wherein the specimen is selected from a nasopharyngeal swab, cerebrospinal fluid, amniotic fluid, serum, plasma, whole blood, bronchopulmonary lavage, nares, vaginal sampling and a rectal/stool sampling obtained from the individual.

12. The rapid method of claim 11, wherein the specimen is a nasopharyngeal swab.

13. The rapid method of claim 7, wherein the antibody is a rabbit polyclonal antibody.

14. The rapid method of claim 7, wherein the HRP-conjugated antibody is an HRP-conjugated anti-rabbit polyclonal goat antibody.

15. The rapid method of claim 7, wherein the binding of the suspected virus SARS-CoV-2 to ACE2-coated microtiter wells is studied in the presence of antibodies contained in convalescent sera or plasma obtained from individuals who have recovered from COVID-19 or from purified monoclonal antibodies.

16. The rapid method of claim 7, wherein the binding of suspected SARS-CoV-2 to ACE2-coated microtiter wells is studied in the presence of drug candidates which may compete for binding and negatively influence the interaction between the viral substrate and its receptor.

17. The rapid method of claim 16, wherein the drug candidates are selected from remdesivir and hydroxychloroquine.

18. A test kit for rapid, highly specific and sensitive, point-of-care detection of a virus in an individual suspected of being infected with a virus, comprising: a plurality of microtiter wells in a microtiter plate, said microtiter wells coated with a host receptor protein specific for a virus deposited on surfaces of the plurality of microtiter wells; an antibody directed against the viral substrate; a wash liquid for washing the plurality of microtiter wells and for preparing a mixture consisting of the wash liquid, an HRP-conjugated antibody directed against the antibody, and a specimen obtained from an individual suspected being infected with the virus, said mixture made into one or more serial dilutions that are deposited atop the coating in the plurality of microtiter wells; a TMB solution; and a STOP solution, wherein the detection of the virus in the specimen is achieved by observing those microtiter wells that undergo a color change, said color change occurring in about five minutes and said detection accomplished by a user in about thirty minutes.

19. The test kit of claim 18, wherein the host receptor protein and the viral substrate that binds to its host receptor protein is selected from SARS-CoV-2:Spike protein and angiotensin-converting enzyme 2 (ACE2), SARS-CoV-2:Spike protein and other host protein candidates, Betacoronaviruses (lineage A):Hemagglutinin (HA) esterase and sialic acid receptors, Influenza:HA protein, sialic acid receptors and HA2; Murine hepatitis virus (MHV):Spike protein and carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1), and Middle East respiratory syndrome (MERS): Spike protein and dipeptidyl peptidase 4 (DPP4/CD26).

20. The test kit of claim 19, wherein the suspected virus is SARS-CoV-2 and the host receptor protein is ACE2.

21. The rapid method of claim 18, wherein the specimen is selected from a nasopharyngeal swab, cerebrospinal fluid, amniotic fluid, serum, plasma, whole blood, bronchopulmonary lavage, nares, vaginal sampling and a rectal/stool sampling obtained from the individual.

22. The test kit of claim 21, wherein the specimen is a nasopharyngeal swab obtained from the individual.

23. The test kit of claim 18, wherein the antibody is a rabbit polyclonal antibody.

24. The test kit of claim 18, wherein the HRP-conjugated antibody is an HRP-conjugated anti-rabbit polyclonal goat antibody.

25. The test kit of claim 18, wherein the infection is COVID-19.

[soupoftheday]

Rapid viral assay

Abstract

The invention provides a method for rapid, highly specific and sensitive detection and quantification of a virus by observing viral substrate binding to its host receptor protein. The invention also provides a method for rapid, highly specific and sensitive detection and quantification of a virus in an individual suspected of being infected with a virus. The invention further provides a test kit for rapid, highly specific and sensitive point-of-care detection of a virus in an individual. The viruses and their host receptor proteins that can rapidly be detected include SARS-CoV-2 and its host receptor protein ACE2. The surprisingly rapid, specific and sensitive method of the invention provides a point-of care test capable of diagnosing individuals suffering from COVID-19 by observation of a color change in the assay, which color change occurs in about five minutes, and which test can be completed by a user in about one hour.

[soupoftheday]

SUMMARY OF THE INVENTION

The present invention fulfills this need by providing a diagnostic method for rapid, highly specific and sensitive, detection and quantification of a virus, such as SARS-CoV-2, which causes COVID-19 disease. The method comprises the steps of coating a plurality of microtiter wells in a microtiter plate with a host receptor protein contained in a coating buffer; incubating the plurality of microtiter wells overnight; washing the microtiter wells; adding a blocking solution to the plurality of microtiter wells; washing the plurality of microtiter wells three times; adding a viral substrate to the plurality of microtiter wells; incubating the plurality of microtiter wells for 20 minutes; washing the plurality of microtiter wells three times; adding an antibody directed against the viral substrate to the plurality of microtiter wells; incubating the plurality of microtiter wells for 20 minutes; adding a horseradish peroxidase (HRP)-conjugated antibody directed against the anti-viral substrate antibody to the plurality of microtiter wells; incubating the plurality of microtiter wells for 20 minutes; washing the plurality of microtiter wells three times; adding a TMB solution to the plurality of microtiter wells; adding a stop solution to the plurality of microtiter wells; and detecting the viral substrate in the microtiter wells by observing those microtiter wells that undergo a color change, or quantifying the concentration of the viral substrate by reading optical density at 450 nm, wherein color change is observed in about five minutes and the method is completed by a user in about one hour.

In another aspect of the present invention, there is provided a diagnostic, point-of-care method for rapid, highly specific and sensitive, detection and quantification of a virus from an individual suspected of being infected with a virus. The method comprises the steps of coating a plurality of microtiter wells with a host receptor protein contained in a coating buffer; incubating the plurality of microtiter wells overnight; washing the microtiter wells; adding a blocking solution to the plurality of microtiter wells; washing the plurality of microtiter wells three times; adding a viral substrate, the viral substrate obtained via a specimen collected from an individual suspected of being infected by a virus or possibly exposed to someone infected with a virus, to the plurality of microtiter wells; incubating the plurality of microtiter wells for 20 minutes; washing the plurality of microtiter wells three times; adding an antibody (i.e., primary antibody) directed against the viral substrate to the plurality of microtiter wells; incubating the plurality of microtiter wells for 20 minutes; adding a horseradish peroxidase (HRP)-conjugated antibody (i.e., secondary antibody) directed against the primary antibody to the plurality of microtiter wells; incubating the plurality of microtiter wells for 20 minutes; washing the plurality of microtiter wells three times; adding a TMB solution to the plurality of microtiter wells; adding a Stop solution to the plurality of microtiter wells; and detecting the viral substrate in the microtiter wells by observing those microtiter wells that undergo a color change, or quantifying the concentration of the viral substrate by reading optical density at 450 nm, wherein the color change is observed about five minutes and the method is completed by a user in about one hour.

In both the above-described methods, after adding a blocking solution to the microtiter wells, the microtiter plate may be stored, after which it can be shipped for use to another site, as the assay start time begins only when the viral substrate is added to the microtiter wells.

The host receptor protein may be, without limitation, ACE2; the viral substrate may be, without limitation, a SARS-CoV-2 Spike protein or a recombinant Spike protein; and the suspected virus may be, without limitation, SARS-CoV-2.

The primary antibody may be, without limitation, a rabbit polyclonal antibody directed against the SARS-CoV-2 Spike protein or the recombinant Spike protein; and the HRP-conjugated antibody directed against the primary antibody may be, without limitation, an HRP-conjugated anti-rabbit polyclonal goat antibody. Tags other than HRP directed against the primary antibody may be used in the invention, including, without limitation, alkaline phosphatase, His, FLAG, or a fluorescent tag. The invention contemplates that any antibodies used, whether they are primary or secondary antibodies, can be either polyclonal or monoclonal, IgG, or IgM, and may be derived from any suitable antibody-producing animal.

The specimen obtained from the individual suspected of being infected by a virus may include, without limitation, a nasopharyngeal swab, cerebrospinal fluid, amniotic fluid, serum, plasma, whole blood, bronchopulmonary lavage, nares, vaginal sampling, semen, or rectal/stool sampling.

In a further aspect of the invention, there is provided a test kit for rapid, highly specific and sensitive, point-of-care detection of a virus from an individual suspected of being infected with the virus. The test kit comprises a plurality of microtiter wells in a microtiter plate, the microtiter wells coated with a host receptor protein specific for the virus deposited on surfaces of the plurality of microtiter wells; a primary antibody directed against the viral substrate; a wash liquid for washing the plurality of microtiter wells and for preparing a mixture consisting of the wash liquid, an HRP-conjugated secondary antibody directed against the primary antibody and a specimen obtained from an individual suspected of being infected with the virus, said mixture made into one or more serial dilutions which are deposited atop the coating in the plurality of microtiter wells; a TMB solution; and a STOP solution, wherein the detection of the virus in the specimen is achieved by observing those microtiter wells that undergo a color change, wherein the color change is observed in about five minutes and the test is completed by a user in about thirty minutes.