Application of Glypican-1 protein in diagnosis of pancreatic cancer, detection method of positive exosome concentration, and use of detection method
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A detection method, exosome technology, applied in the field of biomedicine, can solve the problem of evaluating the value of pancreatic cancer without Glypican-1 protein
Inactive Publication Date: 2017-07-14
AFFILIATED HOSPITAL OF NANTONG UNIV
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[0005] At present, there is no report on the determination of exosome content by nPES in the world, and there is no evaluat...
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Embodiment 1
[0044] Glypican-1 protein is used to prepare products for the diagnosis of pancreatic cancer. Wherein, the product is a chip, a detection reagent or a detection kit.
[0045]Preferably, the chip includes a protein chip, and the detection kit includes a protein immunodetection kit.
Embodiment 2
[0047] A method for detecting the concentration of Glypican-1 positive exosomes is a nanoplasmon enhanced scattering method.
[0048] The detection reagent used in the nano-plasmon-enhanced scattering method includes the following components:
[0050] (II) Phosphate Buffered Washing Solution (PBS), pH 7.4, its components are: 140 mM NaCl, 2.7 mM KCl, 10 mM Na 2 HPO 4 , 1.8mM KH 2 PO 4 ;
[0051] (III) The blocking solution is PBS containing 5% bovine serum albumin (BSA), the pH value is 7.4, and its components are: PBS, 5% BSA;
[0052] (IV) Carboxyl functionalized gold nanoparticles (GNP): 50nm gold nanospheres (radiation green light), 25×60nm gold nanorods (radiation red light), when the two are combined, the signal is enhanced and the yellow light is emitted;
[0053] (V) MES solution, the pH value is 4.7, and its composition is: 2-...
Embodiment 3
[0076] The above-mentioned nanoplasma-enhanced scattering method for detecting the concentration of Glypican-1 positive exosomes was used to detect the content of Glypican-1 protein positive exosomes in the serum of pancreatic cancer for the diagnosis and staging of pancreatic cancer.
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Abstract
The invention discloses an application of a Glypican-1 protein in the diagnosis of pancreatic cancer, a detection method of a Glypican-1 positive exosome concentration, and a use of the detection method. A nano-plasma enhanced scattering (nPES) detection technology can be used to quantify exosomes in serum, an exosome extracting process is omitted, the characteristic antigen of the exosomes is used as a target, and the antigen and a corresponding antibody carrying gold nanoparticles form an antigen-antibody complex in order to obtain exosomes, and the amount of the exosomes is reflected by using the light radiation principle of the gold nanoparticles. The Glypican-1 is a pancreatic cancer-derived exosome biomarker, and the content of Glypican-1 positive exosomes in blood plasma has specific specificity even in early pancreatic cancer, so the exosome content index can be used to diagnose the pancreatic cancer, and the exosome level is closely related to the staging and progression of pancreatic cancer patients. Experiments prove that the method using the nPES detection technology to detect the content of the Glypican-1 exosomes in the serum is concise and accurate, and has better sensitivity and specificity than CA19-9 in the diagnosis of the pancreatic cancer.
Description
technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to the application of a Glypican-1 protein in the diagnosis of pancreatic cancer, a method for detecting the concentration of Glypican-1 positive exosomes and the use thereof. Background technique [0002] Pancreatic cancer is a malignant tumor of the digestive tract with a high degree of malignancy, which is difficult to diagnose and treat. Its morbidity and mortality have increased significantly in recent years. According to statistics from the American Society of Clinical Oncology, the global male pancreatic cancer mortality rate ranks 8th among malignant tumors, and the female patient ranks 9th. The 5-year survival rate of pancreatic cancer patients is always lower than 5%. To make matters worse, pancreatic cancer has no obvious symptoms in the early stage, and the disease course is short and fast. 80% of pancreatic cancer patients live less than 1 year after di...
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