Non-small cell lung cancer detection primer and probe and kit

Abstract

The invention relates to a non-small cell lung cancer detection primer and probe and kit. The kit comprises digital PCR master mix, a droplet stabilizer, miRNAs primer and probe mixed liquid and stem-loop reverse transcription primer mixed liquid, wherein the digital PCR master mix, the droplet stabilizer and the miRNAs primer and probe mixed liquid are used for preparing digital PCR reaction liquid; the stem-loop reverse transcription primer mixed liquid is used for carrying out RNA reverse transcription; the miRNAs primer and probe mixed liquid comprises forward primers and probes and a general reverse primer; the forward primers and probes are used for detecting hsa-mir-125b, hsa-mir-22, hsa-mir-128b and hsa-mir-16; the general reverse primer is used for detecting hsa-mir-125b, hsa-mir-22, hsa-mir-128b and hsa-mir-16; the stem-loop reverse transcription primer mixed liquid comprises stem-loop reverse transcription primers respectively used for reverse transcription of hsa-mir-125b, hsa-mir-22, hsa-mir-128b and hsa-mir-16; hsa-mir-125b, hsa-mir-22 and hsa-mir-128b are markers; hsa-mir-16 is internal reference. The non-small cell lung cancer detection kit can be used for rapidly, conveniently and accurately detecting the miRNA expression levels of non-small cell lung cancer patients.

Description

technical field [0001] The invention relates to a primer probe set and a detection product for detecting the miRNA expression level of non-small cell lung cancer, belonging to the field of biotechnology. Background technique [0002] MicroRNA is a kind of endogenous, highly conserved non-coding small molecule RNA, which is formed by processing single-stranded RNA precursor molecules, and its length is generally 18-23bp. These miRNAs control gene expression at the transcriptional level, post-transcriptional level, and epigenetic level, forming a series of RNA regulatory networks that play an important role in tumorigenesis. Some miRNAs are up-regulated in tumors and act like oncogenes, while some miRNAs are down-regulated in tumors and act as tumor suppressor genes, which have a greater impact on cell functions. Lung cancer is the tumor with the highest morbidity and mortality in the world. The incidence of lung cancer in my country continues to increase every year. More tha...

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Cui et al. detected the expression of miR-125b in the serum of NSCLC patients and found that patients with higher miR-125b expression were less sensitive to cisplatin treatment and had poorer prognosis (Acta Pharmacol Sin, 2013, 34(2)

[0004] Pemetrexed has been widely used in the treatment of advanced non-small cell lung cancer. Among the miRNA markers of non-small cell lung cancer, the expression of miR-22 in serum can be used as a molecular marker for early lung cancer patients. Recent studies have shown that NSCLC patients with higher whole blood miR-22 expression are less sensitive to pemetrexed treatment (2014,229(1):97-99)

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