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Rapid Saliva Tests For SARS-COV-2

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The SARS-COV-2 virus has disrupted global societies in many ways, including direct mortality, indirect mortality from the disruption of health care systems, and the chronic disturbance of global economies(1). SARS-COV-2 is apart of the Coronavirus family and is similar but not drastically the same as other Corona Viruses(2). The SARS-COV-2 has a lipid bilayer in which the N Proteins holds the RNA Genome inside the bilayer, Spike Proteins, which bind to ACE 2 (the receptor expressed in human lungs, arteries, heart, kidney, and intestines(3),) on the outside of the bilayer, together with M and E viral proteins(4). A current possibility for why this new Corona Virus spreads faster is because the new virus's spike can bind to ACE2 10-20 times greater(5).

          The current “gold standard “ for the detection of the virus is the RT-qPCR assay(6). However, even in the most optimal and organized circumstances, this is a slow, laborious assay, and by the time an individual is identified to be positive, they will likely have spread the virus to many other individuals. So, there is a need for a rapid (15 minutes or less), accurate and straightforward assay to enable positive individuals to be quickly isolated. Advanced biomedical technologies for rapid assays have begun to be developed, but the turnaround time of the test is still slow since skilled professionals are needed to obtain the clinical sample (nasopharyngeal swab). This is a slow step andwill be a particular problem in countries where there may be a lack of capable and licensed individuals. In my project, I intend to develop a rapid test using saliva as the test material. This will significantly simplify the sampling process and facilitate the use of the test at work or home by laypeople. I anticipate that I will need generate new very high affinity antibodies to expedite  the rapid assay test.

          It is also imperative to develop simple tests for the presence of anti-COV-SARs-2 antibodies since, at least in the short term, this will enable an immune individual to go about their everyday life. Currently, antibody tests require a blood sample, and again this makes the test complicated, especially in underdeveloped societies. I intend to investigate the presence of anti-covid antibodies in saliva. In preliminary studies, I have been surprised to see that normal saliva contains a high amount of immunoglobulin. I now intend to see whether “normal saliva “ has any Immunoglobulin that recognizes SARS-COV-2 proteins. In particular, I will focus on the coronavirus M protein antigen since I have observed that it is highly similar to the harmless coronavirus OC43, SARS-Cov-1, and SARS-COV-2 (Fig 1). It is possible that these tests will help identify individuals who will be resistant to SARS-COV-2 because of previous exposure to a harmless virus such as Human Corona OC43 and Human Corona HKU1.



Budget

I have been provided with laboratory space and mentorship, and training in the techniques of Biochemistry and Molecular Biology. The prices for these reagents are changing as the market develops, so I will quote rounded approximate estimates for 6 months' work.


Anti-Corona Monoclonal Antibodies - 5,000

Standard immunological reagents (e.g. DAB, secondary antibodies, Colloidal Gold Conjugates,

Avidin-Biotin Peroxidase conjugates) 2500

Custom Cloning of recombinant (corona virus M antigens) - 8000

Custom Synthesis of Antigen peptides -3000

Protein sequencing of Isolated Immunoglobulins -8000



References 


1.Hart, O. E., & Halden, R. U. (2020). Computational analysis of SARS-CoV-2/COVID-19 surveillance by wastewater-based epidemiology locally and globally: Feasibility, economy, opportunities and challenges. Science of The Total Environment, 138875.

2.Hui, D. S., Azhar, E. I., Madani, T. A., Ntoumi, F., Kock, R., Dar, O., ... & Zumla, A. (2020). The continuing 2019-nCoV epidemic threat of novel coronaviruses to global health—The latest 2019 novel coronavirus outbreak in Wuhan, China. International Journal of Infectious Diseases, 91, 264-266.

3.Wu, C., Liu, Y., Yang, Y., Zhang, P., Zhong, W., Wang, Y., ... & Zheng, M. (2020). Analysis of therapeutic targets for SARS-CoV-2 and discovery of potential drugs by computational methods. Acta Pharmaceutica Sinica B.

4.Hamming, I., Timens, W., Bulthuis, M. L. C., Lely, A. T., Navis, G. V., & van Goor, H. (2004). Tissue distribution of ACE2 protein, the functional receptor for SARS coronavirus. A first step in understanding SARS pathogenesis. The Journal of Pathology: A Journal of the Pathological Society of Great Britain and Ireland, 203(2), 631-637.

5.Wrapp, D., Wang, N., Corbett, K. S., Goldsmith, J. A., Hsieh, C. L., Abiona, O., ... & McLellan, J. S. (2020). Cryo-EM structure of the 2019-nCoV spike in the prefusion conformation. Science, 367(6483), 1260-1263.

6.Corman, V. M., Landt, O., Kaiser, M., Molenkamp, R., Meijer, A., Chu, D. K., ... & Mulders, D. G. (2020). Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR. Eurosurveillance, 25(3), 2000045.

Organizer and beneficiary

Will Furneaux
Organizer
Hartford, CT
Julia Furneaux
Beneficiary

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