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Hsa-mir-135a, hsa-mir-302d and hsa-mir-10b joint detection kit for detecting liver cancer genesis or metastasis early

A technology of hsa-mir-302d and hsa-mir-135a, applied in the field of medical biological detection, can solve the joint detection of miR-10b to predict the occurrence and metastasis of liver cancer, and the occurrence or metastasis of miR-302d and cancer. to prevent disease progression and improve prognosis

Inactive Publication Date: 2015-03-11
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] At present, there is no literature report on the relationship between miR-302d and cancer occurrence or metastasis, let alone the application of combined detection of miR-135a, miR-302d and miR-10b to predict the occurrence and metastasis of liver cancer

Method used

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  • Hsa-mir-135a, hsa-mir-302d and hsa-mir-10b joint detection kit for detecting liver cancer genesis or metastasis early
  • Hsa-mir-135a, hsa-mir-302d and hsa-mir-10b joint detection kit for detecting liver cancer genesis or metastasis early
  • Hsa-mir-135a, hsa-mir-302d and hsa-mir-10b joint detection kit for detecting liver cancer genesis or metastasis early

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Embodiment 1: prepare test kit of the present invention (for one person's use)

[0069] The kit of the present invention consists of the following:

[0070]1) Reverse transcription primer (SEQ ID NO:4), 1 tube, concentration: 10 μM, 100 μl / tube;

[0071] 2) Real-time quantitative PCR universal primer (SEQ ID NO:5), 1 tube, concentration: 10 μM, 100 μl / tube;

[0072] miR-135a-specific primer (SEQ ID NO:6), 1 tube, concentration: 10 μM, 100 μl / tube;

[0073] miR-302d specific primer (SEQ ID NO:7), 1 tube, concentration: 10 μM, 100 μl / tube;

[0074] miR-10b specific primer (SEQ ID NO:8), 1 tube, concentration: 10 μM, 100 μl / tube;

[0075] 3) Trizol reagent (total RNA extraction reagent), 1 tube, 2ml / tube

[0076] 4) 1 tube of chloroform, 500 μl / tube;

[0077] 5) 1 tube of isopropanol, 1.500ml / tube

[0078] 6) 1 tube of absolute ethanol, 7ml / tube;

[0079] 7) DEPC H2O (purified water treated with ethylene pyrocarbonate) 1 tube, 1ml / tube

[0080] 8) 1 tube of dd H2O ...

Embodiment 2

[0094] Embodiment 2: the detection method of the kit of embodiment 1

[0095] 1. Serum sample collection and sample preparation:

[0096] Due to the advantages of convenient, non-invasive, and continuous detection of serum, the detection of biomarkers of liver cancer occurrence or metastasis from serum can detect the occurrence or metastasis of liver cancer early, and improve the diagnosis of liver cancer to a new level.

[0097] Plasma samples were collected at Shanghai Oriental Hepatobiliary Surgery Hospital: 40 patients with liver cancer without tumor thrombus, 40 patients with liver cancer with tumor thrombus, and 20 healthy volunteers. Sample collection: Collect about 3-6ml of peripheral blood from the patient and put it into an anticoagulant tube containing 360μl EDTA. Put it into a vacuum coagulation tube, invert it several times to fully combine, let it stand for about half an hour, centrifuge at 1600g for 10min, and transfer the supernatant to a new tube. The supern...

Embodiment 3

[0117] Example 3: Detection of serum hsa-mir-135a / 302d / 10b content in healthy people and patients diagnosed with liver cancer

[0118]The content of hsa-mir-135a / 302d / 10b in the serum of healthy people (20 cases) and liver cancer patients without tumor thrombus (30 cases) were detected respectively according to the detection method of Example 2.

[0119] The result is as figure 1 As shown, the contents of three miRNA molecules hsa-mir-135a / 302d / 10b in the peripheral serum of patients with liver cancer were higher than those of normal patients, in which hsa-mir-135a increased by about 2.5 times, hsa-mir-302d and hsa- The increase of mir-10b was about 2 times, and the difference was statistically significant (P<0.05).

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Abstract

The invention relates to the technical field of biomedical detection, and provides a reagent or a kit for detecting contents of hsa-mir-135a, hsa-mir-302d and hsa-mir-10b in blood plasma or blood serum to predict liver cancer genesis or metastasis as well as a corresponding detection method. The reagent or the kit disclosed by the invention provides a novel approach to diagnose liver cancer genesis and predict live cancer metastasis early, so as to conduct clinical intervention as early as possible, prevent illness development, and improve prognosis for patients.

Description

technical field [0001] The invention relates to the technical field of medical biological detection, and is a kit for detecting the content of hsa-mir-135a, hsa-mir-302d and hsa-mir-10b in plasma or serum to predict the occurrence and metastasis of liver cancer, and its detection method. Background technique [0002] Liver cancer is one of the diseases that seriously threaten people's life and health in our country, and its fatality rate ranks third. About 40% of the patients who die from liver cancer all over the world come from China. With the improvement of treatment methods and imaging detection technology, the five-year survival rate after radical resection of early liver cancer has been greatly improved. However, since most liver cancer patients are already at an advanced stage when they are diagnosed, the overall treatment effect and prognosis of liver cancer patients are relatively poor. [0003] In addition, during the treatment of liver cancer, cancer recurrence...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/118C12Q2600/158C12Q2600/178
Inventor 刘善荣程树群刘淑鹏王录美李明星郭卫星余喜亚徐贵霞
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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