CN114544955A - Application of GASP-2 detection reagent in preparation of lung cancer early diagnosis and susceptibility detection kit - Google Patents
Application of GASP-2 detection reagent in preparation of lung cancer early diagnosis and susceptibility detection kit Download PDFInfo
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Abstract
The invention discloses application of a G protein coupled receptor related sorting protein-2 (GASP-2) detection reagent in preparation of a lung cancer early diagnosis and susceptibility detection kit, belonging to the field of cancer diagnosis. The content of GASP-2 in the serum of the lung cancer patient is obviously higher than that of normal people. The early diagnosis kit for lung cancer can quickly and accurately diagnose the lung cancer by detecting the content of the GASP-2, and is beneficial to improving the early diagnosis rate of the lung cancer, thereby greatly improving the prognosis of lung cancer patients and reducing the disease burden of the lung cancer.
Description
Technical Field
The present invention is in the field of cancer diagnosis.
Background
Lung cancer is the most rapidly growing malignancy of morbidity and mortality worldwide, with over 200 million new lung cancers each year. The lung cancer is the first malignant tumor in morbidity and mortality in China, and the number of newly-released lung cancer is nearly 80 ten thousand and 70 ten thousand in China every year. The incidence rate of lung cancer in China accounts for 37.9 percent of the incidence rate of lung cancer all over the world, and the death rate accounts for 39.4 percent of lung cancer death all over the world. The SEER database in the United states shows that more than half of lung cancer patients are in locally advanced stages or with distant metastases at the time of diagnosis.
At present, the technology for early diagnosis of lung cancer is not mature, and antigens such as CEA (cancer associated tumor markers) and NSE (non-tumor associated tumor markers) which are commonly used clinically at present have limited value in early diagnosis of lung cancer. Therefore, the reagent kit for exploring the early diagnosis of the lung cancer can greatly improve the early diagnosis rate of the lung cancer, thereby greatly improving the prognosis of a lung cancer patient and reducing the disease burden of the lung cancer.
G protein coupled receptor associated sorting protein-2 (G-protein coupled receptor-associated regulatory protein 1, GASP-2, Unit: Q96D09) is a protein that can regulate myogenesis by inhibiting myostatin (myostatin). At present, no report about the relationship between lung cancer and lung cancer exists.
Disclosure of Invention
The invention aims to solve the problems that: provides a new lung cancer early diagnosis kit.
The technical scheme of the invention is as follows:
the application of the reagent for detecting the GASP-2 in preparing a kit for early diagnosis and susceptibility detection of lung cancer (the early diagnosis and the susceptibility detection of the lung cancer can be carried out simultaneously or respectively).
As for the aforementioned use, the reagent for detecting GASP-2 is an ELISA detection reagent.
As for the above-mentioned use, the reagent for detecting GASP-2 is a Western blot reagent.
As used herein, the agent for detecting GASP-2 is a radioimmunoassay agent.
As mentioned above, the reagent for detecting GASP-2 is a protein chip detection reagent.
A kit for early diagnosis of lung cancer, which comprises a reagent for detecting GASP-2.
As with the aforementioned kit, the reagent for detecting GASP-2 is an ELISA detection reagent.
As the kit, the reagent for detecting the GASP-2 is a Western blot reagent.
As with the kit described above, the reagent for detecting GASP-2 is a radioimmunoassay reagent.
As with the kit, the reagent for detecting GASP-2 is a protein chip detection reagent.
The inventor finds that the content of the GASP-2 in the serum of a lung cancer patient (31.55 +/-5.43 ng/ml) is obviously higher than that in the serum of a normal human (4.75 +/-1.26 ng/ml). By detecting the content of the GASP-2 in serum, the lung cancer patient can be effectively identified. If the CUT-OFF value of the serum GASP-2 concentration is defined as: at 12.87ng/ml, the sensitivity of diagnosing lung cancer was 95.1%, and the specificity was 96.0%. The research also finds that the non-lung cancer population with high content of GASP-2 in serum has higher risk of lung cancer.
The kit provided by the invention based on the discovery can quickly and accurately diagnose the lung cancer, screens and identifies the susceptible population of the lung cancer, and is beneficial to improving the early diagnosis rate of the lung cancer, thereby greatly improving the prognosis of lung cancer patients, reducing the disease burden of the lung cancer, and having higher clinical application value.
Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims. In particular, it should be understood that the key point of the present invention is the detection of GASP-2 in serum, and that any method capable of detecting GASP-2 can be used for early diagnosis and susceptibility detection of lung cancer, regardless of the specific means of detection.
The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.
Drawings
FIG. 1 shows the trend of the concentration of GASP-2 in serum between the control group and the group of high risk group. Abscissa unit: and (5) year.
FIG. 2 is a graph showing the trend of the change in serum GASP-2 concentration between the group of persons at high risk of developing lung cancer and the group of persons without lung cancer. Abscissa unit: and (5) year.
Detailed Description
EXAMPLE 1 Lung cancer diagnostic kit (ELISA)
1. Composition of
(1) Pre-coating a plate: ninety-six well plates coated with anti-human GASP-2 rabbit IgG antibody on the inner wall and bottom of the wells.
(2) Enzyme-labeled antibody: (30-fold concentration) HRP-labeled anti-human GASP-2 rabbit IgG antibody.
(3) And (3) standard substance: GASP-2.
(4) Buffer solution: BPS with 1% BSA, 0.05% tween 20.
(5) Color developing agent: TMB substrate solution.
(6) Stopping liquid: 1N sulfuric acid.
2. Use of
(1) Adding 0.1mL of serum sample (standard product) into the pre-coated plate hole, incubating for 30min at 37 ℃, pouring off liquid in the hole, beating for 3 times, and reducing liquid adhesion;
(2) add 0.15mL buffer to wash proteins not bound to pre-coated plate, repeat 5 times;
(3) adding 0.1mL of enzyme-labeled antibody, incubating for 30min at 37 ℃, pouring off the enzyme-labeled antibody, and adding 0.15mL of buffer solution for washing for 5 times;
(4) adding 0.1mL of color developing agent, and incubating for 10min at 37 ℃;
(5) adding 0.05mL of stop solution;
(6) and (4) judging a result: the color depth is observed by naked eyes, and the deeper the color is, the stronger the positive degree is; or measuring the OD (450 nm).
The standard curve can be prepared by the steps of diluting the standard substance with gradient in advance, and the sample dilution degree can be adjusted when the sample is formally detected, so that the concentration falls in a linear interval (a curve of the concentration and the OD value).
The advantageous effects of the present invention are further illustrated in the form of experimental examples.
Experimental example 1 clinical sample testing
1. Method of producing a composite material
Serum samples were collected from 980 lung cancer patients and 1010 normal patients, and the content of GASP-2 in the serum was measured using the kit of example 1.
Wherein: lung cancer patients are 60.8 + -9.0 years old; by gender, male, 624 cases, female: 356 cases; by type, adenocarcinoma: 639, lung adenocarcinoma: 341 example(s). Normal persons are 58.6 ± 7.8 years old; according to gender, male: 612 cases, women: 398 cases; stage I + stage II lung cancer by TNM: 629 cases, stage III + IV lung cancer: 351, the number of cases is 351.
2. Results
The content of the GASP-2 in the serum of a normal human is 4.75 +/-1.26 ng/ml, and the content of the GASP-2 in the serum of a lung cancer patient is 31.55 +/-5.43 ng/ml (p is 0.000).
In lung cancer patients, the content of the GASP-2 in male serum is 33.78 +/-5.43 ng/ml, and the content of the GASP-2 in female serum is 30.87 +/-4.43 ng/ml (p is 0.236); the content of the GASP-2 in the serum of an adenocarcinoma patient is 26.96 plus or minus 2.72ng/ml, the content of the GASP-2 in the serum of a squamous carcinoma patient is 25.19 plus or minus 1.82ng/ml, and the content of the GASP-2 in the serum of a small cell carcinoma patient is 119.94 plus or minus 3.84ng/ml (p is 0.000); the content of the GASP-2 in the blood serum of a stage I lung cancer patient is 14.44 +/-2.42 ng/ml, the content of the GASP-2 in the blood serum of a stage II lung cancer patient is 28.16 +/-1.82 ng/ml, the content of the GASP-2 in the blood serum of a stage III lung cancer patient is 42.78 +/-1.84 ng/ml, and the content of the GASP-2 in the blood serum of a stage IV lung cancer patient is 57.10 +/-3.50/ml (P is 0.000).
In order to discuss the clinical value of detecting the concentration of GASP-2 in serum of a lung cancer patient to diagnose the lung cancer, the diagnostic value of the concentration of GASP2 is calculated by applying a Logistic regression equation in the research, and the result shows that: the sensitivity of detecting the serum GASP-2 concentration of a lung cancer patient to diagnose the lung cancer is 81.59%, the specificity is 86.41%, the positive likelihood ratio is 6.0037, the negative likelihood ratio is 0.2131, the accuracy is 83.58%, the positive predictive value is 89.5%, and the negative predictive value is 76.77% (Table 1).
TABLE 1 logistic regression equation estimation of clinical value for diagnosing lung cancer for detection of GASP-2 concentration
If the CUT-OFF value of the serum GASP-2 concentration is defined as: at 12.87ng/ml, the sensitivity for diagnosing lung cancer was 95.1%, and the specificity was 96.0%, and if the CUT-OFF value at the serum GASP-2 concentration was set to 12.75ng/ml, the sensitivity for diagnosing lung cancer was 90.2%, and the specificity was 96.9%; if the CUT-OFF value of the serum GASP-2 concentration is set to 24.56ng/ml, the sensitivity for diagnosing lung cancer is 95.1%, and the specificity is 99.5%; when the CUT-OFF value was determined to be 30.41ng/ml for the serum GASP-2 concentration, the sensitivity for diagnosing lung cancer was 90.2% and the specificity was 99.5%.
Experimental example 2 tracking of serum GASP-2 concentration and confirmed diagnosis of high risk group of lung cancer
1. Method of producing a composite material
Continuously detecting the serum GASP-2 concentration of high risk group (1233 cases) and control group (899 cases) of lung cancer in high discovery field of lung cancer for 5 years, and performing lung cancer diagnosis on the above groups by using conventional lung cancer diagnosis methods such as medical imaging.
Note:
in the high incidence site of lung cancer, namely, in the area with high incidence rate of lung cancer, Yunnan Xuanwei and personal old are adopted in the experimental example;
the high risk group of lung cancer meets the following conditions: 1. age: is more than or equal to 45 years old; 2. history of smoking: smoking is more than or equal to 20 bags per year; 3. family history of tumor: long term environmental carcinogen exposure.
2. Results
At the end of the 4-year follow-up study period, 128 total populations of the field high risk population group of the high incidence population of lung cancer were diagnosed with lung cancer at different time periods of the study period, and no one of the control population group at the end of the study period was diagnosed with lung cancer; the serum GASP-2 concentration of the control group population has no obvious change in the 4-year research period, while the serum GASP-2 concentration of the high risk group population has a linear rising trend, which shows that the serum GASP-2 concentration gradually rises along with the occurrence and the progress of lung cancer, the comparison of time points between the two groups has significant difference (P is 0.000), and the detection of the serum GASP-2 concentration can over-predict the risk of lung cancer occurrence of the high risk group of lung cancer (fig. 1 and 2).
The results of this experimental example show that: the content of the GASP-2 in serum of high risk population of lung cancer is obviously higher than that of the ordinary population (contrast population), and the susceptibility of lung cancer can be evaluated by detecting the content of the GASP-2 in serum.
In conclusion, the diagnostic kit can assist in diagnosing lung cancer with higher accuracy, and is not limited by the type and stage of lung cancer; and the susceptibility to lung cancer can be detected in advance, and early warning is given to people who do not suffer from lung cancer but have high serum GASP-2 concentration.
Claims (10)
1. The application of the reagent for detecting the G protein-coupled receptor-related sorting protein-2 in preparing a lung cancer early diagnosis and susceptibility detection kit.
2. Use according to claim 1, characterized in that: the reagent for detecting the G protein-coupled receptor related sorting protein-2 is an ELISA detection reagent.
3. Use according to claim 1, characterized in that: the reagent for detecting the G protein-coupled receptor related sorting protein-2 is a Western blot reagent.
4. Use according to claim 1, characterized in that: the reagent for detecting the G protein-coupled receptor-related sorting protein-2 is a radioactive immunoassay reagent.
5. Use according to claim 1, characterized in that: the reagent for detecting the G protein coupled receptor related sorting protein-2 is a protein chip detection reagent.
6. A reagent kit for early diagnosis and susceptibility detection of lung cancer is characterized in that: the kit comprises a reagent for detecting the G protein-coupled receptor-related sorting protein-2.
7. The kit of claim 6, wherein: the reagent for detecting the G protein-coupled receptor related sorting protein-2 is an ELISA detection reagent.
8. The kit of claim 6, wherein: the reagent for detecting the G protein-coupled receptor related sorting protein-2 is a Western blot reagent.
9. The kit of claim 6, wherein: the reagent for detecting the G protein-coupled receptor-related sorting protein-2 is a radioactive immunoassay reagent.
10. The kit of claim 6, wherein: the reagent for detecting the G protein-coupled receptor related sorting protein-2 is a protein chip detection reagent.
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| CN114836542A (en) * | 2022-05-30 | 2022-08-02 | 南方科技大学 | Application of neuroendocrine tumor biomarker in preparation of neuroendocrine tumor diagnostic reagent |
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