阅读说明：本技术 一种乙肝相关肝细胞癌的预后检测试剂盒 (Hepatitis B-related hepatocellular carcinoma prognosis detection kit ) 是由 彭涛 王向坤 朱广志 廖锡文 于 2019-11-11 设计创作，主要内容包括：本发明属于生物医学和临床诊断技术领域,公开了一种乙肝相关肝细胞癌的预后检测试剂盒及其在临床医学中的应用。本发明检测试剂盒中包含载脂蛋白A4、载脂蛋白C3和载脂蛋白C4的检测试剂。具体而言,包含与所述各亚型载脂蛋白相关的免疫吸附剂、酶结合物、酶反应底物和标准品。本发明试剂盒可用于血清、血浆、尿液、胸腹水、灌洗液、脑脊液、组织匀浆等样本的检测。本发明试剂盒由于采用了多种载脂蛋白联合效应分析的策略,使得其对肝细胞癌的预后预测能力得到了显著增强,经验证,本发明试剂盒在预测肝细胞癌的总生存时间和无瘤生存方面灵敏准确、特异性强,另外,本发明试剂盒还具有操作简便、样本易获得、检测耗时短等优点。(The invention belongs to the technical field of biomedicine and clinical diagnosis, and discloses a prognostic detection kit for hepatitis B-related hepatocellular carcinoma and application thereof in clinical medicine. The detection kit comprises detection reagents of apolipoprotein A4, apolipoprotein C3 and apolipoprotein C4. Specifically, the immunopotentiator comprises immunoadsorbent, enzyme conjugate, enzyme reaction substrate and standard substance related to each subtype apolipoprotein. The kit can be used for detecting samples such as serum, plasma, urine, hydrothorax, ascites, lavage fluid, cerebrospinal fluid, tissue homogenate and the like. The kit adopts a strategy of analysis of the combined effect of a plurality of apolipoproteins, so that the prognosis prediction capability of the kit on the hepatocellular carcinoma is obviously enhanced.)

1. A prognosis detection kit for hepatitis B-related hepatocellular carcinoma is characterized in that: the detection kit comprises detection reagents of apolipoprotein A4, apolipoprotein C3 and apolipoprotein C4.

2. The test kit of claim 1, wherein: the detection kit also comprises detection reagents of apolipoprotein A1, apolipoprotein A2, apolipoprotein C1 and apolipoprotein C2.

3. The test kit according to claim 1 or 2, characterized in that: the detection kit comprises immunoadsorbent, enzyme conjugate, enzyme reaction substrate and standard substance which are related to each subtype apolipoprotein.

4. The detection kit according to claim 3, wherein the immunoadsorbent is an antigen or an antibody in a solid phase, the enzyme conjugate is an enzyme-labeled antigen or antibody, and the enzyme reaction substrate is a reaction substrate that undergoes a specific binding reaction with an enzyme in the enzyme conjugate.

5. The test kit of claim 3, wherein: the detection kit also comprises at least one of the following components: sealing liquid, neutral phosphate buffer solution, washing liquid, diluent, color developing agent, stop solution, sodium thiosulfate saturated solution and quality control serum.

6. The test kit according to claim 1 or 2, characterized in that: the detection object of the detection kit is serum, plasma, urine, pleural effusion, lavage fluid, cerebrospinal fluid, cell culture supernatant or tissue homogenate sample.

7. Use of the detection kit according to claim 1 or 2 for the preparation of a diagnostic kit for hepatocellular carcinoma.

8. Use of the detection kit according to claim 1 or 2 for the preparation of a kit for the prognosis evaluation of hepatocellular carcinoma.

Technical Field

The invention belongs to the technical field of biomedicine and clinical diagnosis, and particularly relates to a prognostic detection kit for hepatitis B-related hepatocellular carcinoma and application thereof in clinical medicine.

Background

Liver cancer is ranked as the sixth most common cancer worldwide (Alejandro forter, Llovet, & Bruix,2012), the third most lethal cancer. Among them, hepatocellular carcinoma (HCC) accounts for about 75-80% of primary liver cancer. Risk factors for HCC occurrence include: chronic hepatitis b or c virus (HBV, HCV) infection, type 2 diabetes, heavy drinking, aflatoxin contamination, obesity, non-alcoholic fatty liver disease, and smoking. Today, the treatment of HCC patients relies mainly on surgical resection, local area and systemic treatment, etc. Despite various alternative treatments for HCC, the 5-year survival rate of HCC is still quite low, which is less than 10%, and therefore, it is important to make timely early diagnosis and accurate prognosis evaluation.

Apolipoprotein is a functional protein capable of binding and transporting blood lipids to various tissues of the body for metabolism and utilization, and the Apolipoprotein (APO) family is composed of members of the subfamily APOA, APOB, APOC, APOD, APOE, APOF, APOH, APOL, APOM and APOO according to its different phenotypic classifications. Wherein the subfamilies of apolipoprotein A and apolipoprotein C consist of apolipoprotein A1, apolipoprotein A2, apolipoprotein A4 and apolipoprotein A5, and apolipoprotein C1, apolipoprotein C2, apolipoprotein C3 and apolipoprotein C4, respectively. Studies have found that apolipoprotein family members have various biological activities and are associated with the occurrence and development of various diseases, such as the change of apolipoprotein A1/apolipoprotein B serum ratio is likely to reflect the dynamic balance between atherosclerosis and anti-atherosclerosis, and the ratio is called a good predictor of cerebrovascular and cardiovascular disease risk (Min, Lu, Yong, & Gang, 2011); apolipoprotein C3 is closely related to triglyceride levels (Luo, Su, Yi, Yang, & Peng, 2018); apolipoprotein A1-75G/A and-83C/T genotypes are associated with breast cancer susceptibility and lymph node metastasis (Hamrita,2011) and the like.

However, the research on the APO family focuses mainly on cardiovascular, cerebrovascular and atherosclerosis, and there are few reports on the relation between them and tumors (especially hepatitis b-associated hepatocellular carcinoma), and our research aims to confirm the relation between apolipoprotein and HCC prognosis and to develop a prognostic detection kit for hepatitis b-associated hepatocellular carcinoma based on the relation. To our knowledge, no report on the application of apolipoprotein-based diagnostic techniques to the prognostic assessment of hepatitis b-associated hepatocellular carcinoma has been found, nor has similar detection techniques or detection kits formally been applied in clinical practice.

Disclosure of Invention

In order to carry out accurate prognosis evaluation on hepatitis B related hepatocellular carcinoma (HCC) and search new clinical application for apolipoprotein, a new hepatitis B related hepatocellular carcinoma prognosis detection kit is developed and obtained through a large number of screening and verifying experiments.

The invention relates to a prognosis detection kit for hepatitis B related hepatocellular carcinoma, which is characterized in that: the detection kit comprises detection reagents of apolipoprotein A4, apolipoprotein C3 and apolipoprotein C4.

Further, the detection kit may further comprise detection reagents for apolipoprotein A1, apolipoprotein A2, apolipoprotein C1 and apolipoprotein C2.

Further, the detection kit comprises immunoadsorbent, enzyme conjugate, enzyme reaction substrate and standard substance related to each subtype apolipoprotein.

Further, the immunoadsorbent is an antigen or an antibody in a solid phase, the enzyme conjugate is an antigen or an antibody labeled with an enzyme, and the enzyme reaction substrate is a reaction substrate which performs a specific binding reaction with an enzyme in the enzyme conjugate.

Preferably, the detection kit further comprises at least one of the following components: sealing liquid, neutral phosphate buffer solution, washing liquid, diluent, color developing agent, stop solution, sodium thiosulfate saturated solution and quality control serum.

Preferably, the detection object of the detection kit is serum, plasma, urine, pleural effusion, lavage fluid, cerebrospinal fluid, cell culture supernatant or tissue homogenate sample.

In addition, the invention also relates to the application of the detection kit in the preparation of a hepatocellular carcinoma diagnostic kit.

The invention also relates to application of the detection kit in preparation of a hepatocellular carcinoma prognosis evaluation kit.

In conclusion, the reagent kit adopts a strategy of analyzing the combined effect of a plurality of apolipoproteins, so that the prognosis prediction capability of the reagent kit on the hepatocellular carcinoma is obviously enhanced, and experiments prove that the reagent kit is sensitive, accurate and strong in specificity in the aspects of predicting the total survival time and the tumor-free survival of the hepatocellular carcinoma, and in addition, the reagent kit also has the advantages of simple and convenient operation, easy acquisition of samples, short detection time and the like.

Drawings

FIG. 1 Combined Total survival and Combined tumor-free survival time curves for apolipoprotein A and apolipoprotein C family genes. Joint survival analysis of total survival-related genes: apolipoprotein a4+ C3(a), apolipoprotein a4+ C4(B), apolipoprotein C3+ C4(C), apolipoprotein a4+ C3+ C4 (D); joint survival analysis of tumor-free survival-related genes: apolipoprotein a4+ C4 (E).

FIG. 2 is a total survival risk score model and its survival curve, ROC curve. A: risk score model (risk score ranking, overall survival level, apolipoprotein a4, C3, and C4 expression profile heat map); b: a total survival graph; c: time-dependent ROC curve.

FIG. 3 shows a tumor-free survival risk score model and survival curves and ROC curves thereof. A: risk score model (risk score ranking, tumor-free survival level, apolipoprotein a4 and C4 expression quantity heatmap); b: a tumor-free survival curve graph; c: time-dependent ROC curve.

Detailed Description

The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention.

Before the present embodiments are further described, it is to be understood that the scope of the invention is not limited to the particular embodiments described below; it is also to be understood that the terminology used in the examples is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In addition to the specific methods, devices, and materials used in the examples, any methods, devices, and materials similar or equivalent to those described in the examples may be used in the practice of the invention in addition to the specific methods, devices, and materials used in the examples, in keeping with the knowledge of one skilled in the art and with the description of the invention.

In the present invention, all the equipment and materials are commercially available or commonly used in the industry unless otherwise specified. The methods in the following examples are conventional in the art unless otherwise specified.