Application of protein CTSF (Cathepsin F) in preparation of reagent for diagnosing gastric cancer and diagnostic reagent kit
A diagnostic kit and protein technology, applied in the field of biological sciences, can solve the problems of short survival period of patients, low diagnosis rate of gastric cancer, and operation rate of less than 10%, and achieve the effect of easy operation, high specificity, and auxiliary early diagnosis of gastric cancer.
Active Publication Date: 2014-07-02
SHANGHAI JIAOTONG UNIV
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Problems solved by technology
However, because the vast majority of gastric cancer patients lack specific clinical symptoms, the diagnosis rate of early gastric cancer is very low, and the operation rate is less than 10% (Wu Yunlin: Theory and Practice of Surgery, 2005, 10:401); shorter
[0007] So far, there is no report on the application of cathepsin CTSF in the diagnosis of gastric cancer
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[0037] The present invention is described further in conjunction with embodiment and accompanying drawing.
[0038] 1. Expression, purification and identification of CTSF:
[0039] The protein CTSF is derived from the genetically engineered Saccharomyces cerevisiae in previous studies, induced overexpression by galactose, isolated and purified from agarose affinity medium (glutathione), and identified by Western-Blotting, respectively as follows: figure 1 and figure 2 shown. For a clearer expression, the background with grayscale is used in the figure.
[0040] 2. Preparation of serum samples:
[0041] Centrifuge the whole blood sample at 1000g for about 20 minutes after standing at room temperature for 2 hours or overnight at 4°C, and take the supernatant for immediate detection; or aliquot and store the sample at -20°C or -80°C, but avoid Freeze and thaw repeatedly. Thawed samples should be centrifuged again prior to testing. The tested samples cannot contain NaN 3 ...
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Abstract
The invention discloses application of protein CTSF (Cathepsin F) in preparation of a reagent for diagnosing gastric cancer and a diagnostic reagent kit. An IgG (Immunoglobulin G) antibody for resisting protein CTSF in human serum is qualitatively detected by adopting a traditional clinically-used ELISA (Enzyme-Linked Immunosorbent Assay) technology and applying an indirect method. The method comprises the following specific steps of: coating a microwell plate by using a purified human protein CTSF antigen to obtain a solid-phase antigen; sequentially adding serum to be detected into a micropore coated by the antigen; then adding an ELISA reagent containing an anti-Human IgG antibody labeled by HRP (horse radish peroxidase) to form a CTSF-antibody- enzyme-labelled second antibody complex; after thoroughly washing, adding an enzyme substrate solution TMB (Tetramethylbenzidine) for developing, wherein the TMB is converted into blue under the catalytic action of HRP and is converted into final yellow under the action of acid; and detecting the level of the IgG antibody of the CTSF in the sample by using the color depth. The diagnostic reagent kit provided by the invention is used as a means for assisting in early diagnosis of the gastric cancer, and thus the sensitiveness and the specificity of early diagnosis of the gastric cancer are greatly improved.
Description
technical field [0001] The invention relates to the field of biological sciences, and more specifically relates to the application of protein CTSF in preparing reagents for diagnosing gastric cancer and a diagnostic kit. Background technique [0002] Gastric cancer is a common malignant tumor that endangers human health. According to the statistics of the World Health Organization (WHO) in 2008 (World Health Organization Annual Statistics, 2008), the death rate of gastric cancer patients ranks second in the world. In 2006 WHO statistics showed nearly 950,000 new cases and about 700,000 deaths from this disease. In my country, there are more than 300,000 new gastric cancer patients each year, and about 160,000 people die from gastric cancer. The mortality rate ranks first among malignant tumors, which seriously threatens people's lives and health (Chen Zhu, the third national retrospective sampling survey report on causes of death, 2008) . In order to reduce the mortality ra...
Claims
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IPC IPC(8): G01N33/68
Inventor 杨丽娜陶生策刘炳亚王靖方郭书娟朱正纲李建芳燕敏
Owner SHANGHAI JIAOTONG UNIV