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A probe method digital quantitative PCR kit and its application and usage method

A kit and probe method technology, applied in the field of probe method digital quantitative PCR kits, can solve the problems of difficult identification of lung adenocarcinoma and tuberculous pleurisy, and high detection accuracy, and achieve easy result judgment, high detection efficiency, and high detection efficiency. The effect of improving the efficiency of diagnosis and treatment

Active Publication Date: 2021-10-22
THE WEST CHINA SECOND UNIV HOSPITAL OF SICHUAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The present invention aims to solve the problem that lung adenocarcinoma and tuberculous pleurisy are difficult to distinguish in the prior art, and proposes a probe method digital quantitative PCR kit and its application, which can quickly detect lung adenocarcinoma and tuberculous pleurisy identification, and high detection accuracy

Method used

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  • A probe method digital quantitative PCR kit and its application and usage method
  • A probe method digital quantitative PCR kit and its application and usage method
  • A probe method digital quantitative PCR kit and its application and usage method

Examples

Experimental program
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Effect test

Embodiment 1

[0037] Circular RNA hsa_circ _ 051778 is used as a detection index in the probe method digital quantitative PCR kit, which detects circular RNA hsa_circ in pleural effusion exosome samples _ 051778 copy number as an indicator.

[0038] The above mentioned circular RNA hsa_circ _ 051778 As a detection indicator in the probe method digital quantitative PCR kit, including circular RNA hsa_circ _ 051778 Specific primer, circular RNA hsa_circ _ 051778-specific probe, cDNA obtained after reverse transcription of exosome RNA from pleural effusion of patients with lung adenocarcinoma, and sterile deionized water.

[0039] In this embodiment, the circular RNA hsa_circ _ 051778-specific primers include a front primer and a back primer; the front primer sequence is: 5'-GAAATAAAATGCCAAAGAACACACA-3'; the back primer sequence is 5'-TGGGCAGAGAATGACGAACAG-3'.

[0040] In this embodiment, the circular RNA hsa_circ _ The sequence of the 051778 specific MGB probe is: 5'-TGTCAGCCAGCCAC-3'. ...

Embodiment 2

[0043] The difference between this example and Example 1 is: the circular RNA hsa_circ _ The 5' end of the 051778-specific MGB probe sequence is labeled with the FAM fluorescent reporter group Quencher fluorescence quencher group.

Embodiment 3

[0045] The difference between this example and Example 2 is: the circular RNA hsa_circ _ The 5' end of the 051778-specific MGB probe sequence is labeled with a VIC fluorescent reporter group; the 3' end of the circular RNA hsa_circ_051778-specific MGB probe sequence is labeled with a Minor Groove Binder fluorescence quencher group.

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Abstract

The invention discloses a probe method digital quantitative PCR reagent kit and its application and use method. Circular RNA hsa_circ is disclosed _ 051778 is the application of the detection index in the probe method digital quantitative PCR kit, through the detection of circular RNA hsa_circ in pleural effusion exosome samples _ 051778 The copy number is used as the detection index. The kit includes circular RNA hsa_circ _ 051778 Specific primer, circular RNA hsa_circ _ 051778 specific probe, cDNA obtained after reverse transcription of pleural effusion exosomal RNA from patients with lung adenocarcinoma, PCR amplification master mix for digital PCR by probe method, namely 2x ddPCR Supermix for Probes, and sterile deionized water. Lung adenocarcinoma and tuberculous pleurisy are differentiated by using the detection tool of the present invention, and the detection result shows that the sensitivity of the detection tool can reach more than 90%, which is higher than the single method currently used clinically.

Description

technical field [0001] The invention relates to a kit and its application, in particular to a probe method digital quantitative PCR kit and its application and usage method. Background technique [0002] Lung adenocarcinoma and tuberculous pleurisy are common clinical respiratory diseases. Some clinical manifestations of the two diseases are similar, but their treatment options and prognosis are significantly different. Therefore, accurate diagnosis of these two diseases is very important for the prognosis of patients. important. [0003] Pleural effusion, also known as pleural effusion, refers to the abnormal increase of pleural fluid after the body is affected by certain diseases. Lung adenocarcinoma and tuberculous pleurisy are common causes of pleural effusion, which can be diagnosed by detecting Mycobacterium tuberculosis or tumor cells in pleural effusion, but the positive detection rate is low in actual clinical work, so clinical history and blood tests will also be ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6851C12Q1/6883C12Q1/6886
CPCC12Q1/6851C12Q1/6883C12Q1/6886C12Q2600/158C12Q2600/178C12Q2531/113C12Q2525/307C12Q2563/107
Inventor 王艳云农雪萍张林黄建华车光璐罗海玻周斌
Owner THE WEST CHINA SECOND UNIV HOSPITAL OF SICHUAN
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