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Method for detecting hsa_circ_0007986 as novel biomarker in serum of patient with esophagus cancer and application

A technology of biomarkers and detection methods, applied in the field of tumor molecular biology, can solve the problems of unsatisfactory curative effect, loss of surgical indications, and lack of specific symptoms of esophageal cancer, achieving easy recurrence and metastasis, easy dynamic monitoring, high Effects of sensitivity and specificity

Active Publication Date: 2019-02-22
SUN YAT SEN MEMORIAL HOSPITAL SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, early esophageal cancer lacks specific symptoms, and most patients are diagnosed in the middle and advanced stages, and have lost indications for surgery. Most of them can only rely on systemic chemotherapy and other treatments for disease control, but the curative effect is still unsatisfactory.

Method used

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  • Method for detecting hsa_circ_0007986 as novel biomarker in serum of patient with esophagus cancer and application
  • Method for detecting hsa_circ_0007986 as novel biomarker in serum of patient with esophagus cancer and application
  • Method for detecting hsa_circ_0007986 as novel biomarker in serum of patient with esophagus cancer and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Verification of the differential expression of hsa_circ_0007986 in esophageal squamous patients and healthy subjects

[0024] 1. Collection of serum samples: Each esophageal cancer patient and control group provided 5 mL of fresh blood, collected the upper serum after centrifugation, and stored at -80°C. The controls were all healthy subjects, and the experimental sample information is shown in Table 1 and Table 2.

[0025] Table 1: Basic information of 12 patients with esophageal cancer provided serum

[0026]

[0027] Table 2: Basic information of 12 normal controls

[0028]

[0029]

[0030] 2. For the extraction of serum RNA, refer to the operation procedure in the instruction manual of TRIzol LS Reagent for specific operation steps.

[0031] 3. Reverse transcription: reverse transcription of serum RNA into cDNA, the reverse transcription reaction system is as follows, see Table 3:

[0032] Table 3: Reverse transcription reaction system

[003...

Embodiment 2

[0045] Example 2 Verification of hsa_circ_0007986 as a predictive reagent for esophageal cancer

[0046] 1. Composition of digital PCR kit

[0047] (1) Forward primer: such as SEQ ID NO:2; Reverse primer: such as SEQ ID NO:3;

[0048] (2) Internal reference U6 primer: the forward primer is shown in SEQ ID NO:4; the reverse primer is shown in SEQ ID NO:5;

[0049] (3) For other reagents, refer to SYBR Premix Ex TaqTM II (Tli RNaseH Plus) Quantitative Kit (Code No. RR820A).

[0050] 2. Perform digital PCR detection of hsa_circRNA-0007986 reagent

[0051] (1) Preparation of serum RNA

[0052] Serum samples from another 20 patients with esophageal squamous cell carcinoma were selected, and the specific operation steps of RNA extraction were referred to the operation procedure in the instruction manual of TRIzol LS Reagent. The purity and concentration of the extracted RNA were quantified by NanoDrop ND-1000 Nucleic Acid Quantifier (NanoDrop Technologies, Wilmington, Delaware),...

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Abstract

The invention relates to a method for detecting hsa_circ_0007986 as a novel biomarker in serum of a patient with esophageal cancer. The method mainly comprises digital PCR detection of the biomarker shown as SEQ ID NO:1. Primers used in digital PCR amplification comprise a forward primer shown as SEQ ID NO:2 and a reverse primer shown as SEQ ID NO:3. Hsa_circ_0007986 can be used for preparing a diagnosis reagent and / or drug for esophageal cancer. The method for detecting the esophageal cancer is safe and noninvasive, and even asymptomatic population has high acceptability for the detection; hsa_circ_0007986 is high in accuracy and has higher sensitivity and specificity for the esophageal cancer, is suitable for early screening of the esophageal cancer and serving as the biomarker for prognosis of the esophageal cancer, and cancer relapse and treatment effect are monitored.

Description

technical field [0001] The invention belongs to the field of tumor molecular biology, and in particular relates to a method and application for detecting hsa_circ_0007986 as a biomarker in serum of patients with esophageal cancer. Background technique [0002] Esophageal cancer is the eighth most common malignancy and the sixth most common cause of cancer-related death in the world. Esophageal cancer can be roughly divided into squamous cell carcinoma and adenocarcinoma according to histopathology, and more than 90% of them are squamous cell carcinoma in China. Although the diagnosis and treatment methods related to esophageal cancer have been continuously improved, the prognosis of patients with esophageal cancer is very poor, and the five-year overall survival rate is 15%-34%. At present, the treatment of esophageal cancer is still dominated by surgery, combined with preoperative neoadjuvant chemoradiotherapy or postoperative adjuvant chemoradiotherapy has a certain effec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6851
CPCC12Q1/6851C12Q1/6886C12Q2600/158C12Q2600/166C12Q2600/178C12Q2531/113C12Q2545/101
Inventor 王铭辉胡学廷林华月吴多光王稳健何筱天
Owner SUN YAT SEN MEMORIAL HOSPITAL SUN YAT SEN UNIV
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