Joint detection kit for miRNA marker of lung cancer

A kit and technology for lung cancer, applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of high detection cost, human injury, high false positive of CT, etc., and achieve reliable results, auxiliary screening, and specificity Good results

Inactive Publication Date: 2017-06-09
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, CT false positives are high, detection costs are

Method used

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  • Joint detection kit for miRNA marker of lung cancer
  • Joint detection kit for miRNA marker of lung cancer
  • Joint detection kit for miRNA marker of lung cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1 Detection method of the present invention

[0022] 1. Experimental method

[0023] 1 material

[0024] 1.1 sample

[0025] The tissue samples to be tested were taken from patients admitted to the Department of Respiratory Medicine, West China Hospital of Sichuan University, including 10 lung cancer tissues, 10 normal distal lung cancer tissues, and 10 paracancerous tissues, which have been confirmed by pathology.

[0026] 1.2 Main instruments and reagents

[0027] Bio-Rad CFX96 fluorescent quantitative PCR instrument, analytikjena-scandrop100, Eastep TM Super Total RNA Extraction Kit (Promega), GoScript TM Reverse Transcription System (Promega), Bia-Rad Evagreen mix

[0028] 1.3 Primer design

[0029] Primers were designed according to the miRNA sequence. The detected miRNAs included miR-141 (miRBase Reference Sequence: MIMAT0000432-uaacacugucugguaaagaugg), miR-193b (miRBase Reference Sequence: MIMAT0002819-aacuggcccucaaagucccgcu), and U6 (NCBI Refere...

Embodiment 2

[0079] Embodiment 2 Composition of the detection kit of the present invention and its method of use

[0080] 1. Detection Kit

[0081] 1. The composition of the kit

[0082] Reverse transcription kit (50 copies):

[0083]

[0084] in,

[0085]

[0086] Fluorescent quantitative PCR reaction reverse transcription kit (50 copies):

[0087] components Volume (uL) 2X Evagreen Mix 250ul Forward primer 25ul reverse primer 25ul cDNA 100ul Nuclease-free water 100ul

[0088] in,

[0089]

[0090] 2. How to use the kit

[0091] Tissue samples were crushed by liquid nitrogen grinding, added 300uL tissue lysate and left at room temperature for 5 minutes, then added 300uL diluent and continued to stand at room temperature for 5 minutes, centrifuged at the maximum speed for 5 minutes, carefully sucked up the supernatant, and added 0.5 times the volume of the supernatant in anhydrous ethanol, pipette 3-4 times, and mix well. Transf...

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PUM

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Abstract

The invention discloses a joint detection kit for miRNA marker of lung cancer. The joint detection kit comprises reverse transcription primers shown as SEQ ID NO.1 to 2 as well as primer pairs shown as SEQ ID NO.3 to 4 and SEQ ID NO.5 to 6. The invention also discloses application of the reverse transcription primers shown as SEQ ID NO.1 to 2 as well as the primer pairs shown as SEQ ID NO.3 to 4 and SEQ ID NO.5 to 6 in preparation of reagents for screening the lung cancer. The joint detection kit disclosed by the invention can be used for screening of the lung cancer and clinical auxiliary diagnosis of the lung cancer and has the advantages of high detection sensitivity, good specificity a good clinical application prospect.

Description

technical field [0001] The invention relates to a combined detection kit for miRNA markers of lung cancer. Background technique [0002] Lung cancer is a common malignant tumor, and its morbidity and mortality rate ranks first among other malignant tumors in the world. In the United States in 2015, approximately 86,380 men and 86,380 women died of lung cancer. In China in 2015, the incidence and mortality of lung cancer were 0.0733% and 0.061%, respectively, ranking first among cancer-related diseases. Since the symptoms of early lung cancer patients are not obvious, most patients are already in the middle and late stages when they are diagnosed. Therefore, improving the early diagnosis of lung cancer plays a vital role in the prevention and treatment of lung cancer. At present, the early diagnosis of lung cancer mainly relies on imaging technology and the detection of tumor markers. Low-dose computed tomography (LDCT) has high sensitivity and is the most common means of ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 吴思思朱国念倪银芸丁雨陈雪梅
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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