PATHOGEN SCREENING USING OPTICAL EMISSION SPECTROSCOPY (OES)

Abstract

Apparatus and methods provide discreet and inexpensive screening for pathogens including Covid-19. A sample of bodily fluid such as saliva is energized to generate a plasma, and the optical emission spectra from the plasma is collected and analyzed used a smart optical monitoring system (SOMS) to determine the presence or increase of a protein indicative of a pathogen. The plasma may be generated with a spark, and light may be collected with a smartphone for remote analysis. In particular, in patients with Covid-19 serum concentrations of acute phase proteins (APPs), such as C-reactive protein (CRP) and ferritin, are increased in the cases that develop more severe disease. In addition, increases in serum of several interleukins (IL), such as IL-6 and IL-10, have been described in Covid-19 patients, and these cytokines are known to be mediators of the APPs response.

Claims

1. A method of pathogen screening, comprising the steps of: providing a sample of body fluid; delivering energy to the sample sufficient to generate a plasma; collecting optical emission spectra from the plasma; and analyzing the optical emission spectra to determine the presence or increase of a protein indicative of a pathogen in the body fluid.

2. The method of claim 1, wherein the pathogen is a bacterium, virus, or other microorganism that can cause disease.

3. The method of claim 2, wherein the pathogen is a coronavirus.

4. The method of claim 1, wherein the protein is an acute phase protein (APP) or APP mediator.

5. The method of claim 1, wherein the protein is a C-reactive protein (CRP).

6. The method of claim 1 wherein the protein is ferritin.

7. The method of claim 1 wherein the protein is haptoglobin.

8. The method of claim 1 wherein the protein is amyloid A.

9. The method of claim 1 wherein the protein is an analytes related to an immune response.

10. The method of claim 1 wherein the protein is adenosine deaminase (ADA).

11. The method of claim 1, wherein the protein is an interleukin (IL).

12. The method of claim 10, wherein the interleukin is IL-6 or IL-10.

13. The method of claim 4, wherein the protein is a cytokine.

14. The method of claim 1, wherein the sample of body fluid contains saliva.

15. The method of claim 1, wherein the energy delivered to the sample sufficient to generate a plasma is produced with an electrical spark.

16. The method of claim 1, wherein the optical emission spectra from the plasma is delivered to a smart optical monitoring system (SOMS) via an optical fiber.

17. The method of claim 1, wherein the plasma is using a camera of a smart phone and transmitted as digitized data to a remote SOMS system for analysis.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0011] FIG. 1 shows the use of a SOMS system and spark generator for analysis of a saliva sample;

[0012] FIG. 2 illustrates plasma emission spectroscopy in laser DMD; and

[0013] FIG. 3A presents a result with protein ingestion; and

[0014] FIG. 3B presents a result without protein ingestion.

DETAILED DESCRIPTION OF THE INVENTION

[0015] Optical emission spectroscopy (OES) is a method of chemical analysis that uses the intensity of light emitted from plasma formed during material deposition to determine the quantity and quantity of elements in target objects [3-5]. In addition, the OES collection of the emission spectra generated during an additive manufacturing (AM) process can be used to provide more fundamental physical information, such as the composition of the materials. The emission signature, in addition to chemical composition, can also show the genesis of the spectrum, which can be correlated with various characteristics of the object from which plasma signal is generated.

[0016] As shown in FIG. 1, plasma may be generated by creating a spark where saliva is positioned. The light from the plasma is collected by a lens and transmitted to a SOMS system via fiber optics. The SOMS system breaks down the light into individual spectra, which is sent to a computer to analyze and provide the composition information. It is possible to collect the light using a camera of a smart phone and transmit the digitized data to a central station where the SOMS system and computer are located. Such an arrangement enables an individual to perform the test remotely (i.e., at home), and send the data to a central station for analysis.

[0017] The principle of analysis of the plasma emission is illustrated in FIG. 2. The approach is similar to using a metallic additive manufacturing (AM) process. In AM, metal powders are melted and partially evaporated under the illumination of a highly energetic laser. The metal vapor and shielding gas are excited to high energy level state, with transitions to lower energy level states. In the downward electron transitions, photon wavelengths determined by the energy gaps of the transitions are released and recorded as line-emission spectra. Since the energy gaps are characteristics of elements present, the wavelengths of line emissions in spectra can be used as identifiers for the radiating elements. Further information on emission spectroscopy can be found in [6].

[0018] The spectral image in FIG. 2, for instance, is the spectra collected during laser additive manufacturing of a 7075 aluminum alloy. Peaks at wavelengths, 396.15 nm, 382.94 nm, 357.87 nm, are identified as the peaks of Al, Mg, and Cr [7] which are the main elements in the target material.

[0019] The intensity of the spectrum is proportional to the density of emitted photons. Under the local thermal equilibrium assumption, the emission density (I.sub.i.sub.j (λ)) of photons is:

[00001]$\begin{array}{cc}{I}_{i_j}\left(\lambda \right)=\frac{1}{4\pi }{n}_0{A}_{\mathrm{ij}}\frac{g_i{e}^{-E_i/k_{B}T}}{U\left(T\right)}I\left(\lambda \right)\#.& \left(1\right)\end{array}$

where the partition function U(T) is the statistical occupation fraction of every level k of the atomic species:

U(T)=Σ.sub.jg.sub.je.sup.−E.sup.j.sup./k.sup.B.sup.T#  (2.)

[0020] There are two types of variables associated with this analysis: 1) element-determined variables, including the wavelength of the photon (λ), the transition probability (A.sub.ij), the degeneracy of the upper level (g.sub.i); the energy levels of level i (E.sub.i) and level j (E.sub.j); and 2) the plasma-determined variables, including the number of neutral atoms in plasma (n.sub.0), the temperature of plasma (T), and the spectral line profile I(λ).

[0021] These variables are directly correlated with reference spectra to determine the composition and other properties. For example, the laser power density determines the temperature and electron density of the plasma, which in turn determines the intensity and profile of spectra. Parameters, including laser properties (wavelength, power distribution), powder flow rate, and shielding gas also influence the spectral properties significantly. Therefore, the relationship between spectral signal and manufacturing quality means OES has significant potential for in-situ diagnosis.

Preliminary Results for Protein Identification:

[0022] FIGS. 3A, B show the spectra of saliva with and without protein. An increase in the presence of proteins in saliva can be used to diagnose patients with Covid-19. In particular, in patients with Covid-19 serum concentrations of acute phase proteins (APPs), such as C-reactive protein (CRP) and ferritin, are increased in the cases that develop more severe disease. In addition, increases in serum of several interleukins (IL), such as IL-6 and IL-10, have been described in Covid-19 patients, and these cytokines are known to be mediators of the APPs response.

[0023] Additionally, other APPs such as ferritin, haptoglobin, serum amyloid A, different interleukins, and other analytes related to the immune response, such as adenosine deaminase (ADA), can be measured in saliva. By comparing these proteins between healthy individuals and those with disease, it is possible to assess the differences, which can result from changes in the circulating levels of proteins and/or from changes in the salivary gland secretion, associated with a disease such as Covid-19.

REFERENCES

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