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
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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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