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Application of biomarkers in bile in diagnosis of malignant tumor

A biomarker, bile technology, applied in the application of tools, the field of reagents for detecting biomarkers in bile, can solve the problems of wide application impact, malignant tumor metastasis, lack of verification data, etc., and achieve easy-to-obtain results

Pending Publication Date: 2019-08-16
埃提斯生物技术(上海)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, at present, abdominal X-ray plain film, gallbladder and cholangiography, abdominal B-ultrasound and abdominal CT examination, endoscopic retrograde cholangiopancreatography (ERCP) and percutaneous transhepatic cholangiography (PTC) are mostly used for malignant obstructive jaundice. , Percutaneous Transhepatic Biliary Drainage (PTCD), Percutaneous Transhepatic Biliary Drainage (PTBD) and other methods to diagnose and relieve symptoms, but for the etiology - advanced liver cancer, malignant biliary tumors, Difficult to diagnose and detect features of tumors such as pancreatic cancer
At present, the most direct and effective method is to take tumor tissue by needle biopsy for pathological detection. However, on the one hand, patients with advanced malignant tumors have poor compliance with tissue needle biopsy, and on the other hand, some malignant tumors do have the risk of metastasis. This affects the wide application of needle biopsy in the diagnosis and detection of hepatobiliary and pancreatic malignant tumors; in addition, the specificity of serum tumor markers such as AFP, CEA, and CA19-9 in the diagnosis of liver cancer, malignant biliary tract tumors, and pancreatic cancer Neither the sensitivity nor the sensitivity can meet the clinical needs; moreover, at present, the ctDNA gene mutation detection widely used in lung cancer and other tumors lacks the validation data of its effectiveness in liver cancer, malignant biliary tract tumors, pancreatic cancer, etc.

Method used

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  • Application of biomarkers in bile in diagnosis of malignant tumor
  • Application of biomarkers in bile in diagnosis of malignant tumor
  • Application of biomarkers in bile in diagnosis of malignant tumor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Example 1: Obtaining bile cfDNA

[0079] 1. Sample source:

[0080] 10 BTC patients (4 males, 6 females) were selected, and the specific information is shown in Table 1. The mean age at diagnosis of the 10 BTC patients was 68 years (age range 44-74 years). Four patients had gallbladder cancer and six patients had cholangiocarcinoma (1 intrahepatic, 4 perihilar, and 1 distal). Based on the American Joint Committee on Cancer (AJCC) 7th edition staging system for BTC, 4 patients (40%) were stage II, 3 patients (30%) were stage III, and 3 patients (30%) were stage IV . Percutaneous transhepatic biliary drainage (PTCD) and surgery are considered two different methods of bile extraction. Among the 10 BTC patients, 8 underwent radical resection (R0), and the other 2 underwent R1 and R2 resection.

[0081] Table 1

[0082]

[0083]

[0084] 2. Extract cfDNA:

[0085] A 4-μm-thick paraffin-embedded tissue sample was selected for genomic DNA extraction. DNA was extra...

Embodiment 2

[0086] Example 2: Size distribution of bile cfDNA fragments

[0087] Example 1 Determination of the concentration of extracted bile cfDNA Qubit dsDNA HS Kit (Life Technologies) was used to measure the size and distribution of cfDNA fragments using an Agilent 2100 bioanalyzer (Agilent Technologies, Palo Alto, CA, USA).

[0088] The electropherograms showed that the 10 samples from the above BTC patients all had a similar distribution, which was cut to about 6000bp (eg figure 1 shown). This result suggests that long DNA fragments are more prevalent in bile cfDNA, which is inconsistent with the findings in plasma cfDNA.

Embodiment 3

[0089] Example 3: Mutation of bile cfDNA fragments in BTC patients

[0090] The bile cfDNA extracted in Example 1 was sequenced using the targeted deep sequencing platform (which can simultaneously detect SNVs, indels, and CNVs) of Shanghai 3D Medical Testing Co., Ltd., and a set of 150 tumor-related genes were detected.

[0091] For the evaluation of bile cfDNA samples, unique identifiers were used to capture sequencing, and the targeted deep sequencing platform of Shanghai Roadside Medical Laboratory Co., Ltd. (which can simultaneously detect SNVs, indels, and CNVs) was used to establish variation profiles. 150 tumor-related genes were detected in the bile sample, and the bile cfDNA sample extracted in Example 1 was interrupted with a sonicator (Covaris S2Sonolab, Covaris, Woburn, MA, USA) before sequencing, so that the cfDNA fragments were distributed between 200bp-400bp. The cut-off value of allelic mutation frequency in bile cfDNA samples was ≥0.001.

[0092] The average...

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Abstract

The invention provides an application of reagents for detecting biomarkers in bile in preparation of a tool for diagnosing liver cancer, malignant biliary tract tumors or pancreatic cancer. The invention also provides a tool for diagnosing liver cancer, malignant biliary tract tumors or pancreatic cancer, wherein the tool comprises a reagent for detecting the biomarkers in the bile. The method andtool provided by the invention take the bile as a detection object, has a sample easy to obtain without any adverse effects, and can effectively obtain relevant information involving tumors such as liver cancer, malignant biliary tract tumors or pancreatic cancer.

Description

technical field [0001] The present invention relates to the field of biological detection, and relates to biomarkers present in bile that can be used for diagnosing malignant tumors such as liver cancer, malignant biliary tract tumor, or pancreatic cancer. Specifically, the present invention relates to the preparation of reagents for detecting biomarkers in bile for diagnosis Application in tools for liver cancer, malignant biliary tract tumors or pancreatic cancer. Background technique [0002] Malignant obstructive jaundice is a common type of liver and gallbladder disease, and the degree of damage to the liver and gallbladder is very high. Patients suffering from malignant obstructive jaundice will experience many symptoms, the most important being gallbladder lesions, gallbladder inflammation secretes yellow liquid, commonly known as Jaundice, the harm of jaundice is very serious. The etiology has the following aspects: 1) malignant tumor of the bile duct itself or the ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/156
Inventor 李福根张大东张永杰沈宁佳曹鑫恺周子松
Owner 埃提斯生物技术(上海)有限公司
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