CN108279307B - Blood collection tube for protecting and stabilizing gastrin releasing peptide precursor in serum or plasma - Google Patents
Blood collection tube for protecting and stabilizing gastrin releasing peptide precursor in serum or plasma Download PDFInfo
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- CN108279307B CN108279307B CN201810088563.9A CN201810088563A CN108279307B CN 108279307 B CN108279307 B CN 108279307B CN 201810088563 A CN201810088563 A CN 201810088563A CN 108279307 B CN108279307 B CN 108279307B
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
- G01N33/57423—Specifically defined cancers of lung
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- A—HUMAN NECESSITIES
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- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/15—Devices for taking samples of blood
- A61B5/150007—Details
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- A61B5/15003—Source of blood for venous or arterial blood
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/15—Devices for taking samples of blood
- A61B5/150007—Details
- A61B5/150343—Collection vessels for collecting blood samples from the skin surface, e.g. test tubes, cuvettes
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61B5/150755—Blood sample preparation for further analysis, e.g. by separating blood components or by mixing
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- G—PHYSICS
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
- G01N33/57488—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites involving compounds identifable in body fluids
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Abstract
The invention discloses a blood collection tube with the function of protecting and stabilizing gastrin release peptide precursor in serum or plasma, which comprises a test tube, wherein a sealing rubber plug and a plastic outer cap are arranged at the opening of the test tube, and a protective agent prepared from casein, sodium dodecyl sulfate and aprotinin is pre-filled in the test tube. Because the blood collection tube is pre-filled with the protective agent, the preservation time of the ProGRP sample is greatly prolonged (the sample can be stably preserved for 7 days at the temperature of 2-8 ℃), on one hand, the operation and the application of clinic are convenient, and the retest can be carried out when the detection result of an individual sample is suspected to be abnormal, so that the probability of false negative of the detection result is reduced, and the accuracy and the clinical application value of the detection result of the project are improved.
Description
Technical Field
The invention relates to the field of medical examination, in particular to a blood collection tube for protecting and stabilizing gastrin releasing peptide precursor in serum or plasma.
Background
In recent years, lung cancer has become the first killer of death cause of malignant tumors in China. Lung cancer is classified into non-small cell lung cancer (NSCLC) and Small Cell Lung Cancer (SCLC). Among them, non-small cell lung cancer accounts for about 85%, 5-year survival rate is 15%, and small cell lung cancer accounts for only about 15%, but 2-year survival rate is only 1%. 70% of lung cancer patients are already locally advanced or metastasized at the time of diagnosis, losing the best surgical opportunity. If lung cancer can be diagnosed early, then 5-year survival rates can be improved to over 60%.
Compared with the traditional imaging examination means such as CT, nuclear magnetic resonance and the like, the tumor marker has the advantages of convenience, rapidness, low cost and the like, more importantly, can be helpful for early diagnosis and discovery of cancer, and can be used for assisting in judging the progress, treatment effect and recurrence of the disease through dynamic monitoring of the tumor marker.
Gastrin releasing peptide precursor (ProGRP) is a novel tumor marker for early diagnosis of lung cancer. ProGRP is more advantageous than the former neuron-specific enolase (NSE), especially for small cell lung cancer. The sensitivity of the SCLC for the ProGRP and NSE assays was 73% and 60%, respectively, and the specificity was 98% and 92%, respectively, compared to only 36.84% for SCLC for early diagnosis of NSE, whereas the sensitivity of SCLC for early diagnosis of ProGRP was 63.16%. In addition, ProGRP has a greater advantage over NSE in that the sample test results are not affected by hemolysis.
At present, the detection kits for ProGRP on the market all face the problem of unstable samples when detecting ProGRP. The specification of the ProGRP kit of Yapei corporation: serum samples can only be stored for 3h at 2-8 ℃, and plasma samples can only be stored for 24h at 2-8 ℃. Protocol for ProGRP kit by corneger: serum samples can only be stored for 24h at 2-8 ℃. This is mainly due to the cleavage of ProGRP by proteases in serum. The rapid degradation of the ProGRP sample can cause false negative of the detection result on one hand, and on the other hand, subsequent experiments can not be performed, such as retesting when the detection result of an individual sample is abnormal, which brings great inconvenience to the clinical application of the project.
Disclosure of Invention
In view of the above problems, the present invention provides a blood collection tube for protecting and stabilizing a gastrin-releasing peptide precursor (ProGRP) in serum or plasma; the blood collecting tube can inhibit degradation rate of ProGRP, and prolong storage time of ProGRP sample.
In order to achieve the purpose, the invention can adopt the following technical scheme:
the blood collection tube for protecting and stabilizing the gastrin-releasing peptide precursor in serum or plasma comprises a test tube, wherein a sealing rubber plug and a plastic outer cap are arranged at an opening of the test tube, and a protective agent prepared from casein, sodium dodecyl sulfate and aprotinin is pre-filled in the test tube.
The content of casein in the protective agent is 1-5%, the content of sodium dodecyl sulfate is 1-5%, and the content of aprotinin is 0.1-1 mg/L.
The prepared protective agent has the best effect when the component ratio of casein to sodium dodecyl sulfate to aprotinin is 5:10: 1.
During actual production, the material of the test tube is poly (p-xylylene acid ester), and the material of the sealing rubber plug is butyl rubber.
The vacuum negative pressure of the test tube is 5ml or 10 ml.
When the blood serum or blood plasma centrifugal processing device is used, blood is collected according to a conventional method, the blood is inverted from top to bottom and mixed uniformly, then the blood serum or blood plasma is subjected to centrifugal processing according to a conventional operation method, and the blood serum or blood plasma centrifugal processing device is stored at the temperature of 2-8 ℃.
The invention has the advantages that: because the blood sampling tube is pre-filled with the protective agent, the preservation time of the ProGRP sample is greatly prolonged, on one hand, the method is convenient for clinical operation and application, and on the other hand, retesting can be carried out when the detection result of an individual sample is suspected to be abnormal, so that the probability of false negative of the detection result is reduced, and the accuracy and the clinical application value of the detection result of the project are improved.
Since ProGRP is easily degraded by protease in the sample, the sample storage is unstable. The invention adopts Sodium Dodecyl Sulfate (SDS) (an anionic surfactant) to be matched with SDS with certain concentration, can destroy non-covalent bonds such as ionic bonds, hydrogen bonds and the like in protein, and even change the conformation of the protein, thereby playing a role in blocking the degradation of ProGRP by protease. When the pH of the solution is below the isoelectric point (4.8) of casein, the casein with positively charged molecular surface can be combined with the SDS hydrophilic group with negatively charged by electrostatic interaction to form a casein/SDS complex, and the complex can generate mutual crosslinking action through protein molecules, thereby blocking the degradation of ProGRP by protease.
Drawings
FIG. 1 is a schematic view of the blood collection tube of the present invention.
Detailed Description
As shown in fig. 1, the blood collection tube for protecting and stabilizing gastrin releasing peptide precursor in serum or plasma comprises a test tube 1, a sealing rubber plug 2 and a plastic outer cap 3 are arranged at an opening of the test tube 1, and in order to prolong the storage time of a serum or plasma sample, a protective agent 4 prepared from casein, sodium dodecyl sulfate and aprotinin is prefilled in the test tube 1.
The content of casein in the protective agent is 1-5% (w/v), the content of sodium dodecyl sulfate is 1-5% (w/v), and the content of aprotinin is 0.1-1 mg/L; in actual preparation, the component ratio of casein to sodium dodecyl sulfate to aprotinin in the protective agent 4 is 5:10:1, and the effect is optimal.
The test tube 1 of the invention can be made of hydrophobic poly (p-xylylene acid ester) material, and the sealing rubber plug can be made of butyl rubber.
The vacuum negative pressure of the test tube can be designed to be 5ml or 10ml so as to meet the detection requirement.
EXAMPLE 1 method for producing blood collection tube of the present invention
Firstly, preparing the components of the protective agent 4 (calculated by 1L) in proportion:
1000mL of purified water is measured and sequentially added with NaH2PO4·2H2O 0.59g、Na2HPO4·12H2O5.8 g, NaCl 9g, casein 10g, sodium dodecyl sulfate 20g, aprotinin 2g and ProClin 3001 mL, and stirring until the mixed solution is clear and transparent. Then using a vacuum freeze dryer to carry out freeze-drying treatment to obtain a freeze-dried cake, and then using a homogenizer to treat the freeze-dried cake to obtain uniform powder, namely the protective agent 4.
Secondly, cleaning the inner wall of the test tube 1, and injecting the prepared protective agent 4 into the bottom of the test tube 1 in an ultra-clean bench sterile working environment, wherein the addition amount is 1-5% (w/v) of the blood volume of the blood collection tube;
and thirdly, carrying out vacuum-pumping treatment according to the measurement of the blood collection tube, and then assembling the sealing rubber plug 2 and the plastic outer cap 3, wherein the structure of the finished blood collection tube is shown in figure 1.
Example 2 sample shelf life comparison
The blood collection tube of the present invention and a conventional common tube were used to simultaneously perform blood sample treatment, and 10 samples were examined in total. Turning upside down for 10 times after blood drawing, placing in a 37 deg.C incubator for 30min, taking out, and centrifuging at 4000 rpm for 10 min; and (4) centrifuging and storing at 2-8 ℃. Samples were tested at 0h, 3h, 1 day, 3 days, 5 days, 7 days, and 14 days using a commercially available gastrin-releasing peptide precursor test kit from a company, and the stability of the samples was evaluated under 2-8 ℃ storage conditions for both blood collection tubes, and the results are shown in tables 1 and 2.
Table 12-8 ℃ sample preservation time of blood collection tube of the present invention
Storage time of samples of conventional common blood collection tubes at the temperature of 22-8 ℃ in table
From the experimental data in tables 1 and 2, it can be seen that, by using the blood collection tube with the function of protecting and stabilizing the gastrin-releasing peptide precursor in serum or plasma, the ProGRP sample can be stably stored at 2-8 ℃ for 7 days, and the reduction amplitude is less than 10%. And by using a conventional common tube blood collection tube, the reduction amplitude of the ProGRP sample reaches 20% after the ProGRP sample is stored for 1 day at 2-8 ℃, and the reduction amplitude is larger and larger along with the prolonging of time.
Claims (3)
1. The utility model provides a protection and stable blood collection tube of gastrin release peptide precursor in serum or plasma, includes the test tube opening part is provided with sealed plug and plastics outer cap, its characterized in that: the test tube is pre-filled with a protective agent, and the preparation method of the protective agent comprises the following steps: measuring pureDissolving 1000mL of water, and sequentially adding NaH2PO4·2H2O 0.59g、Na2HPO4·12H2O5.8 g, NaCl 9g, casein 10g, sodium dodecyl sulfate 20g, aprotinin 2g and ProClin 3001 mL, and stirring until the mixed solution is clear and transparent; then using a vacuum freeze drier to carry out freeze-drying treatment to obtain a freeze-dried cake, and then using a homogenizer to treat the freeze-dried cake to obtain a uniform powdery protective agent.
2. The blood collection tube for protecting and stabilizing gastrin-releasing peptide precursor in serum or plasma according to claim 1, wherein: the sealing rubber plug is made of butyl rubber.
3. The blood collection tube for protecting and stabilizing gastrin-releasing peptide precursor in serum or plasma according to claim 1, wherein: the vacuum negative pressure of the test tube is 5ml or 10 ml.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810088563.9A CN108279307B (en) | 2018-01-30 | 2018-01-30 | Blood collection tube for protecting and stabilizing gastrin releasing peptide precursor in serum or plasma |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201810088563.9A CN108279307B (en) | 2018-01-30 | 2018-01-30 | Blood collection tube for protecting and stabilizing gastrin releasing peptide precursor in serum or plasma |
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| CN108279307A CN108279307A (en) | 2018-07-13 |
| CN108279307B true CN108279307B (en) | 2021-01-26 |
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Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109342728A (en) * | 2018-10-18 | 2019-02-15 | 郑州标源生物科技有限公司 | A kind of quality-control product being applicable in tumor markers Pro-GRP quantitative detection |
| CN116718762A (en) * | 2023-06-09 | 2023-09-08 | 上海品峰医疗科技有限公司 | Polypeptide sample stabilizer |
Citations (5)
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| US5770385A (en) * | 1992-06-12 | 1998-06-23 | Tonen Corporation | Antibodies to human gastrin-releasing peptide precursor and use thereof |
| CN102084249A (en) * | 2007-10-26 | 2011-06-01 | 株式会社先端生命科学研究所 | Antibody directed against pro-gastrin releasing peptide, and use thereof |
| CN102101975A (en) * | 2009-12-16 | 2011-06-22 | 罗门哈斯公司 | Low odor compositions and low odor coating compositions |
| CN105158455A (en) * | 2015-09-17 | 2015-12-16 | 深圳市钠科生物有限公司 | Biological sample collecting method, device and system for improving sample detection precision and accuracy, biological sample stabilizing reagent and application |
| CN106701743A (en) * | 2016-12-26 | 2017-05-24 | 广州和实生物技术有限公司 | Blood collection tube for protecting and stabilizing free DNA |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP5201822B2 (en) * | 2006-12-11 | 2013-06-05 | アボット・ラボラトリーズ | Immunological assay for gastrin releasing peptide precursor |
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- 2018-01-30 CN CN201810088563.9A patent/CN108279307B/en active Active
Patent Citations (5)
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|---|---|---|---|---|
| US5770385A (en) * | 1992-06-12 | 1998-06-23 | Tonen Corporation | Antibodies to human gastrin-releasing peptide precursor and use thereof |
| CN102084249A (en) * | 2007-10-26 | 2011-06-01 | 株式会社先端生命科学研究所 | Antibody directed against pro-gastrin releasing peptide, and use thereof |
| CN102101975A (en) * | 2009-12-16 | 2011-06-22 | 罗门哈斯公司 | Low odor compositions and low odor coating compositions |
| CN105158455A (en) * | 2015-09-17 | 2015-12-16 | 深圳市钠科生物有限公司 | Biological sample collecting method, device and system for improving sample detection precision and accuracy, biological sample stabilizing reagent and application |
| CN106701743A (en) * | 2016-12-26 | 2017-05-24 | 广州和实生物技术有限公司 | Blood collection tube for protecting and stabilizing free DNA |
Non-Patent Citations (1)
| Title |
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| 抑肽酶对凝血酶所致星形胶质细胞损伤的保护作用;江承平 等;《中华临床医师杂志（电子版）》;20110131;第5卷(第2期) * |
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