A highly specific spectral detection method for prostate cancer and a cancer judgment device

Abstract

A highly specific spectral detection method for prostate cancer and a cancer judgment device are disclosed. By using a combined detection method of prostate-specific antigen (PSA) and exosomes, the purpose of differentiating the two is achieved through surface plasmon resonance spectroscopy detection, and an application for more simple, rapid, and accurate detection of the serum of prostate cancer patients is realized.

Description

Technical Field

[0001] The present invention relates to the technical fields of plasmonic metasurfaces and biomedical spectral detection, and mainly relates to a method for highly specific spectral detection of prostate cancer and a cancer judgment device. Background Art

[0002] Prostate cancer is one of the main causes of cancer-related death in men and is the most commonly diagnosed cancer in 112 countries. The biomarker prostate-specific antigen (PSA) originates from prostate epithelial cells. It has been widely used in serum tests for the early diagnosis of prostate cancer, which has greatly reduced the mortality rate of prostate cancer. Nevertheless, the low sensitivity and specificity of PSA tests have also led to an increase in overdiagnosis or overtreatment. In particular, an increase in PSA levels in the serum may be closely related to non-malignant diseases, such as benign prostatic hyperplasia (BPH), which usually has symptoms similar to those of prostate cancer. In this case, the diagnostic sensitivity of traditional PSA tests is very low and cannot effectively distinguish prostate cancer, especially in the PSA gray zone (PSA concentration 4 to 10 ng / mL), and invasive prostate biopsy is required for a definite diagnosis. So far, some pioneering efforts have been made to improve the diagnostic accuracy of PSA tests, but for the accurate diagnosis of early prostate cancer, a convenient, non-invasive detection system with high sensitivity and high specificity is still needed.

[0003] Exosomes are extracellular vesicles derived from endosomes, with a size range of 40 nm to 160 nm in diameter. They contain important biological information such as proteins, nucleic acids, lipids, and metabolites from the original cells. In the past few years, exosomes have been associated with the development of human tumors and have attracted interest in cancer diagnosis. Previous studies have shown that a variety of human tumor cells release excessive exosomes, which can be used as in vitro diagnostic biomarkers for cancer to avoid invasive biopsies. So far, researchers have detected the effective diagnosis of exosomes for some cancers, including pancreatic cancer, hepatocellular carcinoma, and lung cancer. However, the detection of exosomes in clinical applications usually faces serious problems such as low speed, low yield, low purity, high cost, and long time consumption, and its use for the diagnosis of prostate cancer has not been reported. Therefore, a simple and efficient serum detection system combining exosomes and PSA should be promising to promote the rapid and accurate diagnosis of prostate cancer. Summary of the Invention

[0004] To solve the above technical problems existing in the prior art, the present invention provides a method for highly specific spectral detection of prostate cancer, including:

[0005] S1: Detect the PSA plasmonic sensor and the exosome plasmonic sensor respectively through spectroscopy to determine the position of the valley of the fundamental PSA resonance wavelength corresponding to the PSA plasmonic sensor and the position of the valley of the fundamental exosome resonance wavelength corresponding to the exosome plasmonic sensor;

[0006] S2: Incubate the PSA plasmonic sensor in known healthy human serum, known tissue hyperplasia patient serum, and known cancer patient serum respectively, and detect the position of the valley of the resonance wavelength through spectroscopy respectively, so as to obtain the first PSA resonance wavelength valley position for known healthy human serum, the second PSA resonance wavelength valley position for known tissue hyperplasia patient serum, and the third PSA resonance wavelength valley position for known cancer patient serum;

[0007] S3: Incubate the exosome plasmonic sensor in known healthy human serum, known tissue hyperplasia patient serum, and known cancer patient serum respectively, and detect the position of the valley of the resonance wavelength through spectroscopy respectively, so as to obtain the first exosome resonance wavelength valley position for known healthy human serum, the second exosome resonance wavelength valley position for known tissue hyperplasia patient serum, and the third exosome resonance wavelength valley position for known cancer patient serum;

[0008] S4: Use the first PSA resonance wavelength valley position, the second PSA resonance wavelength valley position, the third PSA resonance wavelength valley position, and the fundamental PSA resonance wavelength valley position to obtain the first PSA resonance wavelength offset for known healthy human serum, the second PSA resonance wavelength offset for known tissue hyperplasia patient serum, and the third PSA resonance wavelength offset for known cancer patient serum, and use the first exosome resonance wavelength valley position, the second exosome resonance wavelength valley position, the third exosome resonance wavelength valley position, and the fundamental exosome resonance wavelength valley position to obtain the first exosome resonance wavelength offset for known healthy human serum, the second exosome resonance wavelength offset for known tissue hyperplasia patient serum, and the third exosome resonance wavelength offset for known cancer patient serum, and determine the PSA discrimination threshold and the exosome discrimination threshold based on the first, second, and third PSA resonance wavelength offsets and the first, second, and third exosome resonance wavelength offsets.

[0009] Further, before the step S1, there is also a step of biofunctionalizing the PSA plasmonic sensor and the exosome plasmonic sensor for prostate cancer, including:

[0010] Immerse the plasmonic sensor in an ethanol solution for reaction;

[0011] React and clean the plasmonic sensor after the immersion reaction in the ethanol solution with phosphate buffered saline and deionized water to obtain the PSA plasmonic sensor and the exosome plasmonic sensor.

[0012] Further, the first, second, and third PSA resonance wavelength offsets are obtained by performing a difference operation between the valley positions of the first, second, and third PSA resonance wavelengths and the valley position of the baseline PSA resonance wavelength respectively;

[0013] The first, second, and third exosome resonance wavelength offsets are obtained by performing a difference operation between the valley positions of the first, second, and third exosome resonance wavelengths and the valley position of the baseline exosome resonance wavelength respectively.

[0014] Further, the PSA discrimination threshold and the exosome discrimination threshold are obtained based on an ROC analysis of the first, second, and third PSA resonance wavelength offsets and the first, second, and third exosome resonance wavelength offsets.

[0015] Further, the PSA discrimination threshold is the discrimination threshold for distinguishing between healthy individuals and patients with tissue hyperplasia.

[0016] Further, the exosome discrimination threshold includes a first exosome discrimination threshold for distinguishing between healthy individuals and patients with tissue hyperplasia and a second exosome discrimination threshold for distinguishing between patients with tissue hyperplasia and cancer patients.

[0017] Further, the first exosome discrimination threshold is specifically 3.09 nm, and the second exosome discrimination threshold is specifically 5.99 nm.

[0018] The present invention also provides a cancer determination device configured to detect the PSA resonance wavelength offset and the exosome resonance wavelength offset of a test serum using a PSA plasmonic sensor and an exosome plasmonic sensor respectively, and to determine the type of the test serum using the PSA discrimination threshold and the exosome discrimination threshold determined by the method described above.

[0019] Further, the cancer determination device is configured to detect the PSA plasmonic sensor and the exosome plasmonic sensor respectively by spectroscopic detection to determine the valley position of the baseline PSA resonance wavelength of the corresponding PSA plasmonic sensor and the valley position of the baseline exosome resonance wavelength of the corresponding exosome plasmonic sensor;

[0020] The cancer determination device is configured to incubate the PSA plasmonic sensor in the test serum respectively, and to detect the valley position of the resonance wavelength by spectroscopic detection respectively to obtain the valley position of the PSA resonance wavelength for the test serum;

[0021] The cancer determination device is configured to separately place the exosome plasmon sensors into the serum to be tested for incubation, and separately detect the positions of the resonance wavelength valleys through spectral detection to obtain the positions of the resonance wavelength valleys of exosomes in the serum to be tested;

[0022] The cancer determination device is configured to perform a difference operation using the PSA resonance wavelength valley position for the serum to be tested and the basic PSA resonance wavelength valley position to obtain the PSA resonance wavelength offset for the serum to be tested, and perform a difference operation using the exosome resonance wavelength valley position for the serum to be tested and the basic exosome resonance wavelength valley position to obtain the exosome resonance wavelength offset for the serum to be tested.

[0023] Further, the cancer determination device is configured to first determine whether the PSA resonance wavelength offset for the serum to be tested is below the PSA discrimination threshold to determine whether the serum to be tested is serum of a healthy person;

[0024] The cancer determination device is configured to determine whether the exosome resonance wavelength offset for the serum to be tested is below the first exosome discrimination threshold for the serum of healthy persons and patients with tissue hyperplasia to determine whether the serum to be tested is serum of a healthy person;

[0025] The cancer determination device is configured to determine whether the exosome resonance wavelength offset for the serum to be tested is between the first exosome discrimination threshold for the serum of healthy persons and patients with tissue hyperplasia and the second exosome discrimination threshold for the serum of patients with tissue hyperplasia and cancer patients to determine whether the serum to be tested is serum of a patient with tissue hyperplasia;

[0026] The cancer determination device is configured to determine whether the exosome resonance wavelength offset for the serum to be tested is above the second exosome discrimination threshold for the serum of patients with tissue hyperplasia and cancer patients to determine whether the serum to be tested is serum of a cancer patient.

[0027] The technical effect of the present invention is to provide a highly specific spectral detection method for prostate cancer and a cancer determination device. By adopting a combined detection method of prostate cancer specific antigen (PSA) and exosomes, the purpose of distinguishing the two is achieved through plasmon spectral detection, and the application of simpler, faster and more accurate detection of the serum of prostate cancer patients is realized. Description of the Drawings

[0028] The accompanying drawings are included to provide a further understanding of the embodiments and are incorporated in and constitute a part of this specification. The drawings illustrate the embodiments and, together with the description, are used to explain the principles of the invention. Other embodiments and many of the intended advantages of the embodiments will be readily apparent as they become better understood by reference to the following detailed description. The elements of the drawings are not necessarily to scale with each other. Like reference numerals refer to corresponding like parts.

[0029] Figure 1 The flowchart of a high-specificity spectral detection method for prostate cancer according to an embodiment of the present invention is shown;

[0030] Figure 2 The scanning electron microscope image of the surface of a plasmonic sensor according to a specific embodiment of the present invention is shown;

[0031] Figure 3 The 45° side-view scanning electron microscope image of a plasmonic sensor according to a specific embodiment of the present invention is shown;

[0032] Figure 4 The comparison diagram of experimental results of a PSA plasmonic sensor for detecting PSA in the sera of known healthy people, patients with benign prostatic hyperplasia, and cancer patients according to a specific embodiment of the present invention is shown;

[0033] Figure 5 The scanning electron microscope image after a plasmonic sensor for exosomes detects exosomes in the sera of known healthy people, patients with benign prostatic hyperplasia, and cancer patients according to a specific embodiment of the present invention is shown;

[0034] Figure 6 The comparison diagram of experimental results obtained by nanoparticle tracking analysis of exosomes and by the method of using a plasmonic sensor for exosomes in the sera of known healthy people, patients with benign prostatic hyperplasia, and cancer patients according to a specific embodiment of the present invention is shown;

[0035] Figure 7 The comparison diagram of experimental results of a plasmonic sensor for exosomes for detecting CD63 in the sera of healthy people, patients with benign prostatic hyperplasia, and cancer patients according to a specific embodiment of the present invention is shown;

[0036] Figure 8 The resonance wavelength shift distribution diagram for distinguishing 10 healthy people and 20 patients with benign prostatic hyperplasia obtained by the threshold value from ROC according to a specific embodiment of the present invention is shown;

[0037] Figure 9 The resonance wavelength shift distribution diagram for distinguishing 10 healthy people and 13 patients with benign prostatic hyperplasia in the PSA gray zone obtained by the threshold value from ROC according to a specific embodiment of the present invention is shown;

[0038] Figure 10 Shows the resonance wavelength shift distribution diagram for distinguishing 20 patients with benign prostatic hyperplasia and 20 patients with prostate cancer by the threshold obtained through ROC according to a specific embodiment of the present invention;

[0039] Figure 11 Shows the wavelength shift obtained by distinguishing the sera of 13 PSA gray zone patients among 20 patients with benign prostatic hyperplasia and the sera of 13 PSA gray zone patients among 20 patients with prostate cancer by the threshold obtained through ROC according to a specific embodiment of the present invention;

[0040] Figure 12 Shows the ROC curve graph of the resonance wavelengths of sera from 20 patients with benign prostatic hyperplasia and 20 patients with prostate cancer obtained by the method of PSA concentration and PSA-bound exosomes according to a specific embodiment of the present invention;

[0041] Figure 13 Shows the ROC curve graph of the resonance wavelengths of sera from 13 patients with benign prostatic hyperplasia and 13 patients with prostate cancer in the PSA gray zone obtained by the method of PSA concentration, serum free PSA / total PSA (F / T-PSA), and PSA-bound exosomes according to a specific embodiment of the present invention;

[0042] Figure 14 Shows the comparison graph of the sensitivity, specificity, positive predictive value, negative predictive value, accuracy of prostate cancer in the PSA gray zone and magnetic resonance after PSA detection according to a specific embodiment of the present invention. Detailed implementation manners

[0043] The present invention will be further described in detail below with reference to the drawings and embodiments. It can be understood that the specific embodiments described herein are only used to explain the related invention, rather than limiting the invention. Additionally, it should be noted that for the sake of description, only parts related to the relevant invention are shown in the drawings.

[0044] It should be noted that, without conflict, the embodiments in the present invention and the features in the embodiments can be combined with each other. The present invention will be described in detail below with reference to the drawings and embodiments.

[0045] According to an embodiment of the present invention, a highly specific spectral detection method for prostate cancer is provided.

[0046] Figure 1 Shows the highly specific spectral detection method for prostate cancer according to an embodiment of the present invention, as Figure 1 shown, the method includes:

[0047] S1: Detect the PSA plasmonic sensor and exosome plasmonic sensor respectively through spectral detection to determine the position of the trough of the fundamental PSA resonance wavelength corresponding to the PSA plasmonic sensor and the position of the trough of the fundamental exosome resonance wavelength corresponding to the exosome plasmonic sensor.

[0048] Further, before the step S1, there is also a step of biofunctionalizing the PSA plasmonic sensor and the exosome plasmonic sensor for prostate cancer, including:

[0049] Immerse the plasmonic sensor in an ethanol solution for reaction. The surface and side-view scanning electron microscope images of the plasmonic sensor are as Figure 2 and Figure 3 shown. This step specifically includes immersing the plasmonic sensor in an ethanol solution of MUA (1 mM) at 4 °C for 12 hours to form a self-assembled layer;

[0050] React and clean the plasmonic sensor after soaking reaction in the ethanol solution based on phosphate buffered saline and deionized water to obtain the PSA plasmonic sensor and the exosome plasmonic sensor. This step specifically includes immersing the plasmonic sensor in a mixture of 400 mM EDC and 100 mM NHS in phosphate buffered saline (PBS) for 60 minutes to activate the terminal carboxyl groups of MUA. Wash all the plasmonic sensors with deionized water, divide them into two groups, and incubate them with 100 μg / mL anti-PSA in PBS and 12.5 μg / mL anti-CD63 in PBS for 30 minutes respectively. Next, place all the plasmonic sensors in 50 μg / mL BSA in PBS for 30 minutes to block unbound anti-PSA or anti-CD63, and rinse with deionized water to obtain the PSA plasmonic sensor and the exosome plasmonic sensor for subsequent biosensing.

[0051] S2: Place the PSA plasmonic sensors into known healthy human serum, known tissue hyperplasia patient serum, and known cancer patient serum respectively for incubation, and detect the position of the trough of the resonance wavelength through spectral detection respectively to obtain the first PSA resonance wavelength trough position for known healthy human serum, the second PSA resonance wavelength trough position for known tissue hyperplasia patient serum, and the third PSA resonance wavelength trough position for known cancer patient serum.

[0052] Further, the first, second, and third PSA resonance wavelength offsets are obtained by performing difference operations on the positions of the troughs of the first, second, and third PSA resonance wavelengths respectively and the position of the trough of the fundamental PSA resonance wavelength.

[0053] Furthermore, the serum of the known tissue hyperplasia patient is the serum of the benign prostatic hyperplasia patient, and the serum of the known cancer patient is the serum of the prostate cancer patient.

[0054] Preferably, the PSA plasmonic sensor tested the sera of 10 known healthy individuals, 20 sera of known benign prostatic hyperplasia patients, and 20 sera of known prostate cancer patients, respectively, generating resonance wavelength shifts in the ranges below 0.19 nm, 0.31 - 0.82 nm, and 0.38 - 2.02 nm. Among them, 13 PSA gray zone patients included in the 20 sera of known benign prostatic hyperplasia patients and 13 PSA gray zone patients included in the 20 sera of known prostate cancer patients generated resonance wavelength shifts in the ranges of 0.31 - 0.63 nm and 0.38 - 0.57 nm, respectively.

[0055] Furthermore, as Figure 4 shown, a comparison graph of the experimental results of the PSA plasmonic sensor for detecting PSA in the sera of known healthy individuals, benign prostatic hyperplasia patients, and cancer patients according to a specific embodiment of the present invention is shown. The vertical axis is the resonance wavelength shift. It can be seen that the distribution of the resonance wavelength shifts of the sera of benign prostatic hyperplasia patients and prostate cancer patients has no obvious pattern. Among them, **** indicates that when the significance level is 0.0001, the means of the resonance wavelength shifts of the corresponding sera are significantly different, and ns indicates that there is no correlation between the two groups of samples. From the experimental results, it can be seen that there is no obvious difference in the resonance wavelength shifts of the sera of benign prostatic hyperplasia patients and prostate cancer patients detected by the PSA plasmonic sensor, and it is impossible to directly distinguish the types of sera of benign prostatic hyperplasia patients and prostate cancer patients by detecting PSA in the sera.

[0056] S3: Place the exosome plasmonic sensor into the sera of known healthy individuals, known tissue hyperplasia patients, and known cancer patients for incubation, and detect the positions of the resonance wavelength valleys through spectral detection respectively to obtain the first exosome resonance wavelength valley position for the sera of known healthy individuals, the second exosome resonance wavelength valley position for the sera of known tissue hyperplasia patients, and the third exosome resonance wavelength valley position for the sera of known cancer patients.

[0057] Preferably, the exosome plasmonic sensor and the PSA plasmonic sensor are placed into the sera of known healthy individuals, known tissue hyperplasia patients, and known cancer patients for incubation respectively, and the incubation time is specifically 20 - 30 minutes.

[0058] Furthermore, the first, second, and third exosome resonance wavelength shifts are obtained by performing difference operations on the positions of the first, second, and third exosome resonance wavelength valleys and the position of the basic exosome resonance wavelength valley respectively.

[0059] Preferably, the exosome plasmon sensor was tested with 10 sera from known healthy individuals, 20 sera from known patients with benign prostatic hyperplasia, and 20 sera from known patients with prostate cancer, generating resonance wavelength shifts in the ranges below 2.21 nm, 3.97 - 6.06 nm, and 5.80 - 8.26 nm, respectively. Among them, 13 patients in the PSA gray zone among the 20 sera from known patients with benign prostatic hyperplasia and 13 patients in the PSA gray zone among the 20 sera from known patients with prostate cancer generated resonance wavelength shifts in the ranges of 3.97 nm - 6.06 nm and 5.93 nm - 8.26 nm, respectively.

[0060] Further, as Figure 5 shown, a scanning electron micrograph of the exosome plasmon sensor according to a specific embodiment of the present invention for detecting exosomes in sera of known healthy individuals, patients with benign prostatic hyperplasia, and cancer patients is shown. From left to right are the SEM images of the bare chip, healthy individuals, patients with benign prostatic hyperplasia, and patients with prostate cancer after incubation and binding with exosomes. The oval or circular particles with diameters between 40 - 160 nm in the figure are exosomes. It can be seen from the figure that there are significant differences in the number of exosomes in the serum samples of patients with benign prostatic hyperplasia and prostate cancer within the same area. As Figure 6 shown, a comparison graph of the experimental results obtained by nanoparticle tracking analysis of exosomes and the method using the exosome plasmon sensor in sera of known healthy individuals, patients with benign prostatic hyperplasia, and cancer patients according to a specific embodiment of the present invention is shown. The left y-axis coordinate in this figure corresponds to the resonance wavelength shifts obtained by the exosome plasmon sensor in sera of known healthy individuals, known patients with benign prostatic hyperplasia, and known patients with prostate cancer, and the right y-axis coordinate corresponds to the concentration results of exosomes obtained by nanoparticle tracking analysis of exosomes. It can be seen from the figure that the results obtained by the two methods show a positive correlation. Combining Figure 5 and Figure 6 proves the feasibility of the scheme for differentiating sera of patients with benign prostatic hyperplasia and prostate cancer through exosomes.

[0061] Further, as Figure 7As shown, a comparison graph of experimental results of exosome plasmon sensors according to a specific embodiment of the present invention for detecting CD63 in the sera of healthy individuals, patients with benign prostatic hyperplasia, and cancer patients is presented. The vertical axis represents the resonance wavelength shift. Among them, **** indicates that when the significance level is 0.0001, the mean values of the resonance wavelength shifts are all significantly different. It can be seen that including the sera of healthy individuals, the distribution of the resonance wavelength shifts of sera from different populations is significantly regular. By performing a two-sample t-test on the results, it is found that when the significance level is 0.0001, the mean values of the resonance wavelength shifts are all significantly different, indicating that there are obvious differences in the resonance wavelength shifts of clinical serum samples from prostate cancer patients and patients with benign prostatic hyperplasia. By combining Figure 5 , Figure 6 and Figure 7 it is further illustrated that the sera of patients with benign prostatic hyperplasia and prostate cancer patients can be distinguished by exosomes.

[0062] S4: Obtain a first PSA resonance wavelength shift for known healthy human serum, a second PSA resonance wavelength shift for known serum of patients with tissue hyperplasia, and a third PSA resonance wavelength shift for known cancer patient serum by using the first PSA resonance wavelength trough position, the second PSA resonance wavelength trough position, the third PSA resonance wavelength trough position, and the basic PSA resonance wavelength trough position, and obtain a first exosome resonance wavelength shift for known healthy human serum, a second exosome resonance wavelength shift for known serum of patients with tissue hyperplasia, and a third exosome resonance wavelength shift for known cancer patient serum by using the first exosome resonance wavelength trough position, the second exosome resonance wavelength trough position, the third exosome resonance wavelength trough position, and the basic exosome resonance wavelength trough position, and determine a PSA discrimination threshold and an exosome discrimination threshold based on the first, second, and third PSA resonance wavelength shifts and the first, second, and third exosome resonance wavelength shifts.

[0063] Furthermore, the PSA discrimination threshold and the exosome discrimination threshold are obtained based on a ROC analysis of the first, second, and third PSA resonance wavelength shifts and the first, second, and third exosome resonance wavelength shifts.

[0064] Furthermore, the PSA discrimination threshold is the discrimination threshold for the sera of healthy individuals and patients with tissue hyperplasia. By using the PSA plasmonic sensor to test the sera of 10 known healthy individuals, 20 known patients with prostate hyperplasia, and 20 known patients with prostate cancer respectively, resonance wavelength shifts in the ranges below 0.19 nm, 0.31 - 0.82 nm, and 0.38 - 2.02 nm are generated. Among the 20 known patients with prostate hyperplasia, 13 patients in the PSA gray zone and among the 20 known patients with prostate cancer, 13 patients in the PSA gray zone generate resonance wavelength shifts in the ranges 0.31 - 0.63 nm and 0.38 - 0.57 nm respectively. The PSA discrimination threshold adopts the discrimination threshold of 0.31 nm between the sera of patients with prostate hyperplasia and healthy individuals.

[0065] Furthermore, the exosome discrimination threshold includes a first exosome discrimination threshold for the sera of healthy individuals and patients with tissue hyperplasia and a second exosome discrimination threshold for the sera of patients with tissue hyperplasia and cancer patients.

[0066] Preferably, as Figure 8 shown, the wavelength shifts obtained from the sera of 10 known healthy individuals and 20 known patients with prostate hyperplasia are subjected to receiver operating characteristic curve (ROC) analysis, and the threshold for discriminating between healthy individuals and patients with prostate hyperplasia is 3.09 nm.

[0067] Preferably, as Figure 9 shown, the wavelength shifts obtained from the sera of 10 known healthy individuals and 13 patients in the PSA gray zone among 20 known patients with prostate hyperplasia are subjected to ROC analysis, and the threshold for discriminating between the sera of healthy individuals and patients with prostate hyperplasia is 3.09 nm.

[0068] Preferably, as Figure 10 shown, the wavelength shifts obtained from the sera of 20 known patients with prostate hyperplasia and 20 known patients with prostate cancer are subjected to ROC analysis, and the threshold for discriminating between the sera of patients with prostate hyperplasia and prostate cancer is 5.99 nm.

[0069] Preferably, as Figure 11 shown, the wavelength shifts obtained from the sera of 13 patients in the PSA gray zone among 20 known patients with prostate hyperplasia and 13 patients in the PSA gray zone among 20 known patients with prostate cancer are subjected to ROC analysis, and the threshold for discriminating between the sera of patients with prostate hyperplasia and prostate cancer is 5.99 nm. Through Figure 8 、 Figure 9 、 Figure 10 and Figure 11 the experimental results, the specific value of the first exosome discrimination threshold is 3.09 nm, and the specific value of the second exosome discrimination threshold is 5.99 nm.

[0070] Further, as Figure 12 shown, the ROC curve graph of the resonance wavelengths of the sera of 20 patients with benign prostatic hyperplasia and 20 patients with prostate cancer obtained by the PSA concentration and the method of PSA-bound exosomes is shown. Among them, the area under the ROC curve (AUC) obtained by the method of PSA or exosomes is 99.0%, which is much higher than the AUC value of 61.8% obtained only by the PSA concentration method. The experimental results illustrate the high sensitivity of the method adopted by the present invention in sera in the PSA gray zone and above the PSA gray zone.

[0071] Further, as Figure 13 shown, the ROC curve graph of the resonance wavelengths of the sera of 13 patients with benign prostatic hyperplasia and 13 patients with prostate cancer in the PSA gray zone obtained by the PSA concentration, serum free PSA / total PSA (F / T-PSA), and the method of PSA-bound exosomes is shown. Among them, the area under the ROC curve (AUC) obtained by the method of PSA-bound exosomes is 99.4%, which is much higher than the AUC value of 61.8% obtained only by the PSA concentration method and the value of 75.1% obtained by the F / T-PSA method. The experimental results illustrate the high sensitivity of the method adopted by the present invention in sera in the PSA gray zone.

[0072] To highlight the sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of the proposed method for prostate cancer in the PSA gray zone, the method of the present invention is compared with magnetic resonance imaging after PSA detection. As Figure 14 shown, the sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of prostate cancer detection obtained by magnetic resonance imaging after PSA detection are 23.1%, 92.3%, 75.0%, 54.5%, and 57.7% respectively. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of prostate cancer obtained by the present invention are all 92.3%, all higher than those of the magnetic resonance imaging method after PSA detection.

[0073] According to an embodiment of the present invention, a cancer determination device is provided.

[0074] The cancer determination device is configured to respectively detect the PSA resonance wavelength shift amount and the exosome resonance wavelength shift amount of the serum to be tested by using a PSA plasmonic sensor and an exosome plasmonic sensor, and determine the type of the serum to be tested by using the PSA discrimination threshold and the exosome discrimination threshold determined by the method.

[0075] Further, the cancer determination device is configured to detect a PSA plasmon sensor and an exosome plasmon sensor respectively through spectral detection to determine the position of the base PSA resonance wavelength valley corresponding to the PSA plasmon sensor and the position of the base exosome resonance wavelength valley corresponding to the exosome plasmon sensor;

[0076] The cancer determination device is configured to incubate the PSA plasmon sensors respectively in the serum to be tested, and detect the position of the resonance wavelength valley respectively through spectral detection to obtain the PSA resonance wavelength valley position for the serum to be tested;

[0077] The cancer determination device is configured to incubate the exosome plasmon sensors respectively in the serum to be tested, and detect the position of the resonance wavelength valley respectively through spectral detection to obtain the exosome resonance wavelength valley position for the serum to be tested;

[0078] The cancer determination device is configured to perform a difference operation using the PSA resonance wavelength valley position for the serum to be tested and the base PSA resonance wavelength valley position to obtain the PSA resonance wavelength offset for the serum to be tested, and perform a difference operation using the exosome resonance wavelength valley position for the serum to be tested and the base exosome resonance wavelength valley position to obtain the exosome resonance wavelength offset for the serum to be tested.

[0079] Preferably, when incubating the exosome plasmon sensors and the PSA plasmon sensors respectively in the serum to be tested, the incubation time is specifically 20 - 30 minutes.

[0080] Further, the cancer determination device is configured to first determine whether the PSA resonance wavelength offset for the serum to be tested is below the PSA discrimination threshold to determine whether the serum to be tested is from a healthy person;

[0081] The cancer determination device is configured to determine whether the exosome resonance wavelength offset for the serum to be tested is below the first exosome discrimination threshold for the sera of healthy persons and patients with tissue hyperplasia to determine whether the serum to be tested is from a healthy person;

[0082] The cancer determination device is configured to determine whether the exosome resonance wavelength offset for the serum to be tested is between the first exosome discrimination threshold for the sera of healthy persons and patients with tissue hyperplasia and the second exosome discrimination threshold for the sera of patients with tissue hyperplasia and cancer patients to determine whether the serum to be tested is from a patient with tissue hyperplasia;

[0083] The cancer determination device is configured to determine whether the exosome resonance wavelength shift amount for the serum to be tested is above a second exosome discrimination threshold for serum of tissue hyperplasia patients and cancer patients, so as to determine whether the serum to be tested is the serum of a cancer patient.

[0084] The specific embodiments of the present invention have been described above, but the protection scope of the present invention is not limited thereto. Any person skilled in the art within the technical scope disclosed by the present invention can easily think of changes or substitutions, which should all be covered within the protection scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.

[0085] In the description of the present invention, it should be understood that the orientation or positional relationship indicated by the terms "upper", "lower", "inner", "outer", etc. is based on the orientation or positional relationship shown in the drawings. It is only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation. Therefore, it should not be construed as a limitation to the present invention. The word 'comprising' does not exclude the presence of elements or steps not listed in the claims. The word 'a' or 'an' preceding an element does not exclude the presence of a plurality of such elements. The simple fact that certain measures are recited in mutually different dependent claims does not indicate that a combination of these measures cannot be used to advantage. Any reference signs in the claims should not be construed as limiting the scope.

Claims

1. A highly specific spectral detection method for prostate cancer, characterized in that, Including: S1: Detect the PSA plasmon sensor and exosome plasmon sensor respectively through spectral detection to determine the position of the trough of the basic PSA resonance wavelength corresponding to the PSA plasmon sensor and the position of the trough of the basic exosome resonance wavelength corresponding to the exosome plasmon sensor; S2: Place the PSA plasmon sensors into known healthy human serum, known tissue hyperplasia patient serum, and known cancer patient serum respectively for incubation, and detect the positions of the troughs of the resonance wavelengths respectively through spectral detection to obtain the first PSA resonance wavelength trough position for known healthy human serum, the second PSA resonance wavelength trough position for known tissue hyperplasia patient serum, and the third PSA resonance wavelength trough position for known cancer patient serum; S3: Place the exosome plasmon sensors into known healthy human serum, known tissue hyperplasia patient serum, and known cancer patient serum respectively for incubation, and detect the positions of the troughs of the resonance wavelengths respectively through spectral detection to obtain the first exosome resonance wavelength trough position for known healthy human serum, the second exosome resonance wavelength trough position for known tissue hyperplasia patient serum, and the third exosome resonance wavelength trough position for known cancer patient serum; S4: Use the first PSA resonance wavelength trough position, the second PSA resonance wavelength trough position, the third PSA resonance wavelength trough position, and the basic PSA resonance wavelength trough position to obtain the first PSA resonance wavelength offset for known healthy human serum, the second PSA resonance wavelength offset for known tissue hyperplasia patient serum, and the third PSA resonance wavelength offset for known cancer patient serum, and use the first exosome resonance wavelength trough position, the second exosome resonance wavelength trough position, the third exosome resonance wavelength trough position, and the basic exosome resonance wavelength trough position to obtain the first exosome resonance wavelength offset for known healthy human serum, the second exosome resonance wavelength offset for known tissue hyperplasia patient serum, and the third exosome resonance wavelength offset for known cancer patient serum, and determine the PSA discrimination threshold and exosome discrimination threshold based on the first, second, and third PSA resonance wavelength offsets and the first, second, and third exosome resonance wavelength offsets.

2. The method according to claim 1, characterized in that, Before the step S1, there is also a step of biofunctionalizing the PSA plasmon sensor and the exosome plasmon sensor for prostate cancer, including: Immerse the plasmon sensor in an ethanol solution for reaction; React and clean the plasmon sensor after soaking reaction in the ethanol solution based on phosphate buffered saline and deionized water to obtain the PSA plasmon sensor and the exosome plasmon sensor.

3. The method according to claim 2, characterized in that, The first, second, and third PSA resonance wavelength offsets are obtained by performing difference operations on the first, second, and third PSA resonance wavelength trough positions respectively and the basic PSA resonance wavelength trough position; The first, second, and third exosome resonance wavelength offsets are obtained by performing difference operations on the trough positions of the first, second, and third exosome resonance wavelengths and the trough position of the basal exosome resonance wavelength, respectively.

4. The method according to claim 3, characterized in that, The PSA discrimination threshold and the exosome discrimination threshold are obtained based on ROC analysis of the first, second, and third PSA resonance wavelength offsets and the first, second, and third exosome resonance wavelength offsets.

5. The method according to claim 4, characterized in that, The PSA discrimination threshold is the discrimination threshold for the sera of healthy individuals and patients with tissue hyperplasia.

6. The method according to claim 4, characterized in that, The exosome discrimination threshold includes a first exosome discrimination threshold for the sera of healthy individuals and patients with tissue hyperplasia and a second exosome discrimination threshold for the sera of patients with tissue hyperplasia and cancer patients.

7. The method according to claim 6, characterized in that, The specific value of the first exosome discrimination threshold is 3.09 nm, and the specific value of the second exosome discrimination threshold is 5.99 nm.

8. A cancer judgment device: configured to detect the PSA resonance wavelength shift and exosome resonance wavelength shift of the serum to be tested by using a PSA plasmon sensor and an exosome plasmon sensor respectively, and judge the type of the serum to be tested by using the PSA discrimination threshold and exosome discrimination threshold determined by the method according to any one of claims 1-7.

9. The device according to claim 8, characterized in that, The cancer determination device is configured to detect a PSA plasmonic sensor and an exosome plasmonic sensor respectively through spectral detection to determine the trough position of the basal PSA resonance wavelength of the corresponding PSA plasmonic sensor and the trough position of the basal exosome resonance wavelength of the corresponding exosome plasmonic sensor; The cancer determination device is configured to incubate the PSA plasmonic sensors respectively in the serum to be tested and detect the trough positions of the resonance wavelengths respectively through spectral detection to obtain the trough positions of the PSA resonance wavelengths for the serum to be tested; The cancer determination device is configured to incubate the exosome plasmonic sensors respectively in the serum to be tested and detect the trough positions of the resonance wavelengths respectively through spectral detection to obtain the trough positions of the exosome resonance wavelengths for the serum to be tested; The cancer determination device is configured to perform a difference operation on the trough position of the PSA resonance wavelength for the serum to be tested and the trough position of the basal PSA resonance wavelength to obtain the PSA resonance wavelength offset for the serum to be tested, and perform a difference operation on the trough position of the exosome resonance wavelength for the serum to be tested and the trough position of the basal exosome resonance wavelength to obtain the exosome resonance wavelength offset for the serum to be tested.

10. The device according to claim 9, characterized in that, The cancer determination device is configured to first determine whether the PSA resonance wavelength offset for the serum to be tested is below the PSA discrimination threshold to determine whether the serum to be tested is the serum of a healthy individual; The cancer determination device is configured to determine whether the exosome resonance wavelength offset for the serum to be tested is below the first exosome discrimination threshold for the sera of healthy individuals and patients with tissue hyperplasia to determine whether the serum to be tested is the serum of a healthy individual; The cancer determination device is configured to determine whether the exosome resonance wavelength offset for the serum to be tested is between the first exosome discrimination threshold for the sera of healthy individuals and patients with tissue hyperplasia and the second exosome discrimination threshold for the sera of patients with tissue hyperplasia and cancer patients to determine whether the serum to be tested is the serum of a patient with tissue hyperplasia; The cancer determination device is configured to determine whether the exosome resonance wavelength shift amount for the serum to be tested is above a second exosome discrimination threshold for sera of tissue hyperplasia patients and cancer patients, so as to determine whether the serum to be tested is the serum of a cancer patient.

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