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A kind of dye method digital quantitative PCR kit and its application and use method

A kit and copy number technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of increasing the economic burden of patients, low positive detection rate, cumbersome identification process, etc., to achieve easy result judgment and detection. The effect of high throughput and improved diagnosis and treatment efficiency

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

AI Technical Summary

Problems solved by technology

Lung adenocarcinoma and tuberculous pleurisy are common causes of pleural effusion. At present, the diagnosis can be made by detecting Mycobacterium tuberculosis or tumor cells in pleural effusion, but the diagnosis will be confirmed clinically through medical history, blood test, pleural effusion biochemical test, and erythrocyte sedimentation rate test. , tuberculosis antibody detection, cell chromosome detection, tumor antigen detection, etc. to make differential diagnosis between the two, the identification process is cumbersome, time-consuming, and increases the economic burden of patients, so the positive detection rate in actual clinical work is low

Method used

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  • A kind of dye method digital quantitative PCR kit and its application and use method
  • A kind of dye method digital quantitative PCR kit and its application and use method

Examples

Experimental program
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Effect test

Embodiment 1

[0043]The application of circular RNA hsa_circ_0051778 as a detection indicator in the preparation of a dye-based digital quantitative PCR kit for the identification of lung adenocarcinoma and tuberculous pleurisy, by detecting the copy number of circular RNA hsa_circ_0051778 in pleural effusion exosome samples as a detection indicator, if pleural effusion If the copy number of circular RNA hsa_circ_0051778 in the exosome sample is greater than 4.1 copies / well, it is judged that the patient is a tuberculous pleurisy patient, otherwise, the patient is a lung adenocarcinoma patient.

Embodiment 2

[0045] A dye method digital quantitative PCR reagent kit includes specific primers, positive control, negative control and PCR amplification premix, wherein the negative control is sterile deionized water.

Embodiment 3

[0047] On the basis of Example 2, further: the specific primer is a circular RNA hsa_circ_0051778 specific primer, including a front primer sequence of 5'-GATCTCTCCGTTCTCAGGCC-3', and a back primer sequence of 5'-GAGAATGACGAACAGGAGCG-3';

[0048] The positive control is cDNA reverse-transcribed from RNA in pleural effusion exosomes of patients with lung adenocarcinoma;

[0049] The PCR amplification master mix is ​​QX200 ddPCR EvaGreen Supermix.

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Abstract

The invention discloses a dye method digital quantitative PCR kit and its application and usage method. Including the application of the circular RNA hsa_circ_0051778 copy number as a detection indicator in the differentiation kit between lung adenocarcinoma and tuberculous pleurisy, according to the copy number of circular RNA hsa_circ_0051778 in tuberculous pleurisy is higher than that in lung adenocarcinoma; Among them, the kit includes circular RNA hsa_circ_0051778-specific primers, positive control, negative control, PCR amplification premix for dye-based digital PCR and sterile deionized water; through sampling, PCR amplification system preparation, amplification reaction and identification Steps to use this kit. Therefore, it can accurately and quickly identify lung adenocarcinoma and tuberculous pleurisy. The detection results show that the sensitivity of this 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 dye method digital quantitative PCR kit and its application and use method, in particular to a dye method digital quantitative PCR kit based on circular RNA hsa_circ_0051778 as a detection index and its application and use method, belonging to medical detection tools . Background technique [0002] Lung adenocarcinoma and tuberculous pleurisy are both clinically common respiratory diseases, and 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 of great importance to patients. Prognosis is extremely important. [0003] Pleural effusion refers to the abnormal increase of pleural fluid caused by certain diseases affecting the pleura. Lung adenocarcinoma and tuberculous pleurisy are common causes of pleural effusion. At present, the diagnosis can be made by detecting Mycobacterium tuberculosi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12Q1/6883C12Q1/6851
CPCC12Q1/6851C12Q1/6883C12Q1/6886C12Q2600/178C12Q2531/113C12Q2537/16C12Q2545/113
Inventor 王艳云张林车光璐农雪萍黄建华罗海玻周斌
Owner THE WEST CHINA SECOND UNIV HOSPITAL OF SICHUAN
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