Portable diagnostic device for viewing biological entities and structures

Abstract

The present invention seeks to provide an easily portable means of visualizing biological samples by way of fluorescent emissions from fluorescently tagged antibodies associated with the biological entities; thereby acting as a diagnostic tool when appropriate antibodies and samples are applied to the sample plate. Current methods used to visualize biological entities by way of fluorescent emissions from fluorescently tagged antibodies associated with the biological samples involve the use of large bulky equipment that doesn't exist in a modular format—different components existing as disparate disjointed units that cannot be physically associated or linked with each other. This invention significantly decreases the size of the components needed to visualize biological samples by way of fluorescent emissions from fluorescently tagged antibodies and also modularizes the components such that they can be connected to each other to form the portable detection device.

Claims

1. A method of visualizing biological entities or structures using a single portable device, comprising: (a) directly or indirectly applying a biological sample that is directly or indirectly associated with a fluorescently-tagged antibody onto a sample plate; (b) physically connecting the sample plate with contained fluorescently-tagged biological sample to an excitation-wavelength-source support structure that holds light emitting diodes that emit excitation wavelengths of appropriate wavelength targeted at the fluorescently-tagged antibody associated with the biological sample (light emitting diodes being powered by a power source connected to the portable device) such that the sample plate and excitation-wavelength-source support structure form one continuous object; (c) amplifying and focusing the emitted excitation wavelength from the fluorescently-tagged antibody associated with the biological entity towards a detection device using a stack of appropriate lenses or single lens physically connected to the sample plate and excitation-wavelength-source support structure so that the sample plate, excitation-wavelength-source support structure and lens(es) form a single modular structure.

Description

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWING

[0013] FIG. 1 is a side view of the device showing excitation of the fluorophore bound to the antibody and fluorescent emission following excitation.

[0014] FIG. 2 is the top view of the excitation-wavelength-source showing the arrangement of the excitation-wavelength-source light emitting diodes.

[0015] FIG. 3 is a longitudinal cross-section of the excitation-wavelength-source support structure showing the arrangement of the inner and outer walls of the excitation-wavelength-source support structure and the transmission channel for the electromagnetic energy released by the excitation-wavelength-source light emitting diode.

DETAILED DESCRIPTION OF THE INVENTION

[0016] FIG. 1a Is a side view of the detection device. The detection device will have the capacity to capture emitted excitation wavelengths from the fluorescent tag (FIG. 1k) bound to antibody (FIG. 1j) associated with the target of interest. Such detection device will also have the capacity to record and store captured images of the emitted excitation wavelengths.

[0017] FIG. 1b shows a representation of a fluorescent emission from an excited fluorophore (FIG. 1k). The fluorescent emission will result from the absorption of electromagnetic energy directed from an excitation-wavelength-source light emitting diode (FIG. 1d).

[0018] FIG. 1c shows a representation of the magnifying-plane stack. The magnifying-plane stack amplifies and focuses the emitted excitation wavelength (FIG. 1b) emitted by an excited fluorophore (FIG. 1k).

[0019] FIG. 1d shows one excitation-wavelength-source light emitting diode. The excitation-wavelength-source light emitting diodes are contained in the excitation-wavelength source support structure (FIG. 1m) and they act to release electromagnetic energy at a specific wavelength tuned to excite a given fluorophore.

[0020] FIG. 1e shows excitation wavelength travelling to target fluorophore.

[0021] FIG. 1f shows biological entity bound to sample plate wall. The biological entity possesses an antigen that is recognizable by the fluorescently tagged antibody (FIG. 1j).

[0022] FIG. 1g shows the exterior wall of the sample plate.

[0023] FIG. 1h shows the power source for the excitation-wavelength-source light emitting diode.

[0024] FIG. 1i shows the exterior wall of the enclosure of the power source for the excitation-wavelength-source light emitting diode.

[0025] FIG. 1j shows primary antibody with fluorescent tag attached to target surface. Attachment of the primary antibody to the target surface is dependent on recognition of a specific epitope.

[0026] FIG. 1k shows fluorophore emitting fluorescence following excitation by excitation wavelength.

[0027] FIGS. 1l and 1o show the lower and upper portals respectively of the excitation-wavelength-source support structure through which fluorescent emissions from the fluorophore (FIG. 1k) traverse prior to capture by the detection device (FIG. 1a). The lower and upper excitation portals are created by the inner wall (FIG. 1p) of the excitation-wavelength-source support structure and they minimize contamination of the fluorescent emission (FIG. 1b) from the fluorescent tag with excitation wavelength released by the excitation-wavelength-source light emitting diode (FIG. 1d).

[0028] FIG. 1m shows the excitation-wavelength-source exterior wall support structure.

[0029] FIG. 1n shows the excitation-wavelength-source support structure inter-wall connector.

[0030] FIG. 1p shows the inner wall of the excitation-wavelength-source support structure.

[0031] FIG. 2a shows a top view of the exterior wall of the excitation-wavelength-source support structure.

[0032] FIG. 2b shows a top view of the excitation-wavelength-source light emitting diode.

[0033] FIG. 2c shows a top view of the inner wall of the excitation-wavelength-source support structure.

[0034] FIG. 2d shows a top view of the upper portal of the excitation-wavelength-source support structure.

[0035] FIG. 2e. shows a top view of the excitation-wavelength-source support structure inter-wall connector.

[0036] FIG. 3a shows a longitudinal view of the excitation-wavelength-source light emitting diode.

[0037] FIG. 3b. is a representation of an excitation wavelength from the excitation-wavelength-source light emitting diode traversing the transmission channel of the excitation-wavelength-source support structure.

[0038] FIG. 3c. shows a longitudinal view of the inner wall of the excitation-wavelength-source support structure.

[0039] FIG. 3d. shows a longitudinal view of the outer wall of the excitation-wavelength-source support structure.

[0040] FIG. 3e. shows a longitudinal view of the transmission channel of the excitation-wavelength-source support structure.

[0041] FIG. 3I shows the point of contact between the vertical section of the exterior wall of the excitation-wavelength-source support structure and the lower circular portion of the exterior wall of the excitation-wavelength-source support structure.

[0042] FIG. 3II shows the point of contact between the vertical section of the interior wall of the excitation-wavelength-source support structure and the lower circular portion of the interior wall of the excitation-wavelength-source support structure.

BIBLIOGRAPHY

[0043] 1. Weiss, S. R. & Leibowitz, J. L. Coronavirus Pathogenesis. in Advances in Virus Research vol. 81 85-164 (Elsevier, 2011). [0044] 2. Wang, H. et al. The genetic sequence, origin, and diagnosis of SARS-CoV-2. Eur J Clin Microbiol Infect Dis 39, 1629-1635 (2020). [0045] 3. Wu, A. et al. Genome Composition and Divergence of the Novel Coronavirus (2019-nCoV) Originating in China. Cell Host & Microbe 27, 325-328 (2020). [0046] 4. He, Y. et al. Receptor-binding domain of SARS-CoV spike protein induces highly potent neutralizing antibodies: implication for developing subunit vaccine. Biochemical and Biophysical Research Communications 324, 773-781 (2004). [0047] 5. Xu, X. et al. Evolution of the novel coronavirus from the ongoing Wuhan outbreak and modeling of its spike protein for risk of human transmission. Sci. China Life Sci. 63, 457-460 (2020). [0048] 6. Hu, B. et al. Discovery of a rich gene pool of bat SARS-related coronaviruses provides new insights into the origin of SARS coronavirus. PLoS Pathog 13, e1006698 (2017). [0049] 7. Konrad, R. et al. Rapid establishment of laboratory diagnostics for the novel coronavirus SARS-CoV-2 in Bavaria, Germany, February 2020. Eurosurveillance 25, (2020). [0050] 8. Corman, V. M. et al. Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR. Eurosurveillance 25, (2020). [0051] 9. Chan, J. F.-W. et al. Improved Molecular Diagnosis of COVID-19 by the Novel, Highly Sensitive and Specific COVID-19-RdRp/Hel Real-Time Reverse Transcription-PCR Assay Validated In Vitro and with Clinical Specimens. J Clin Microbiol 58, e00310-20, /jcm/58/5/JCM.00310-20.atom (2020). [0052] 10. Reusken, C. B. E. M. et al. Laboratory readiness and response for novel coronavirus (2019-nCoV) in expert laboratories in 30 EU/EEA countries, January 2020. Eurosurveillance 25, (2020). [0053] 11. Pan, Y., Zhang, D., Yang, P., Poon, L. L. M. & Wang, Q. Viral load of SARS-CoV-2 in clinical samples. The Lancet Infectious Diseases 20, 411-412 (2020). [0054] 12. Wei, S. et al. Field-deployable, rapid diagnostic testing of saliva samples for SARS-CoV-2. http://medrxiv.org/lookup/doi/10.1101/2020.06.13.20129841 (2020) doi:10.1101/2020.06.13.20129841. [0055] 13. Harikrishnan, P. Saliva as a Potential Diagnostic Specimen for COVID-19 Testing. J Craniofac Surg (2020) doi:10.1097/SCS.0000000000006724. [0056] 14. Loeffelholz, M. J. et al. Multicenter Evaluation of the Cepheid Xpert Xpress SARS-CoV-2 Test. J Clin Microbiol 58, e00926-20, /jcm/58/8/JCM.00926-20.atom (2020). [0057] 15. Chau, C. H., Strope, J. D. & Figg, W. D. COVID-19 Clinical Diagnostics and Testing Technology. Pharmacotherapy phar.2439 (2020) doi:10.1002/phar.2439. [0058] 16. Baek, Y. H. et al. Development of a reverse transcription-loop-mediated isothermal amplification as a rapid early-detection method for novel SARS-CoV-2. Emerging Microbes & Infections 9, 998-1007 (2020). [0059] 17. Broughton, J. P. et al. CRISPR-Cas12-based detection of SARS-CoV-2. Nat Biotechnol 38, 870-874 (2020). [0060] 18. Sandstrom, E. et al. Detection of human anti-HTLV-III antibodies by indirect immunofluorescence using fixed cells. Transfusion 25, 308-312 (1985). [0061] 19. Hedenskog, M. et al. Testing for antibodies to AIDS-associated retrovirus (HTLV-III/LAV) by indirect fixed cell immunofluorescence: Specificity, sensitivity, and applications. J. Med. Virol. 19, 325-334 (1986). [0062] 20. Simonetti, R. F., Dewar, R. & Maldarelli, F. Diagnosis of Human Immunodeficiency Virus Infection. in Principles and Practive of Infectious Diseases vol. 1 (Elsevier, 2015).