Composition for noninvasive screening of early lung cancer, application, kit and sample treatment method

A composition and kit technology, applied in the field of compositions for non-invasive screening of early lung cancer, can solve the problems of insufficient detection sensitivity, lack of markers for early detection and prognosis evaluation of lung cancer, etc.

Inactive Publication Date: 2020-10-23
上海纳奥生物科技有限公司
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Problems solved by technology

However, the currently marketed lung cancer methylation detection kits (SHOX-2, RASSF1A) are limited to tumor tissue level and alveolar lavage fluid samples due to insufficient detection sensitivity.
However, markers for early detection and prognosis evaluation of lung cancer with high sensitivity and specificity through patient plasma are still lacking.

Method used

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  • Composition for noninvasive screening of early lung cancer, application, kit and sample treatment method

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Experimental program
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Effect test

Embodiment 1

[0035] Detection of methylation of CDH1 and SHP-1 genes in plasma cell-free DNA of patients with stage I lung cancer. The detection reagents for specifically detecting the DNA methylation of the human CDH1 gene are primers and probes for the human CDH1 gene; The sequence modified by bisulfite or hydrazine salt; more preferably, the detection reagent for specifically detecting the DNA methylation of human CDH1 gene is: shown in SEQ ID NO: 1 and SEQ ID NO: 2 Primers and the probe shown in SEQ ID NO: 3; or the detection reagent for specifically detecting the DNA methylation of the human SHP-1 gene is a primer and a probe for the SHP-1 gene; preferably, the The primers and probes are directed at the sequence modified by bisulfite or bisulfite or hydrazine salt in the human SHP-1 gene; more preferably, the DNA methyl group of the specific detection human SHP-1 gene The detection reagents used are: the primers shown in SEQ ID NO: 4 and SEQ ID NO: 5 and the probe shown in SEQ ID NO:...

Embodiment 2

[0060] The plasma samples from 200 cases of lung cancer and 100 cases of healthy people from Shanghai Pulmonary Hospital were tested for the methylation of CDH-1 and SHP-1 genes in these 300 cases of plasma samples. In the test results of 100 normal plasma samples, the difference between the Ct values ​​of CDH-1 and SHP-1 genes and the Ct values ​​of internal reference genes was greater than 8, and the specificity reached 95%. The difference between the Ct value of the target gene and the Ct value of the internal reference gene in 192 samples was less than 8, and the sensitivity reached 97%, including 92% for stage 0 lung cancer and 98% for stage I-II lung cancer.

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Abstract

The invention relates to a composition for noninvasive screening of early lung cancer, application, a kit and a sample treatment method. The composition comprises a detection reagent for specific detection of DNA methylation of a human CDH1 gene; and a detection reagent for specifically detecting DNA methylation of the human SHP-1 gene. The kit for detecting methylation sites of the CDH1 gene andthe SHP-1 gene not only comprises a promoter region of the gene, but also comprises a coding region and a non-coding region of the gene, so that the target sequences of the CDH1 gene and the SHP-1 gene which are highly related to the early lung cancer can be found through multi-region detection, meanwhile, the detection accuracy and specificity can be improved, and the detection rate of methylation of the CDH1 gene and the SHP-1 gene is increased. The kit has strong pertinence on early screening of lung cancer at the molecular level, and has the advantages of good detection specificity, high sensitivity, good reliability and the like, so that early possible lung cancer patients can be prevented as soon as possible, and corresponding treatment measures can be taken.

Description

technical field [0001] The present invention relates to the technical field of early screening of lung cancer, more specifically, the present invention relates to a composition, application, kit and sample processing method for non-invasive screening of early lung cancer. Background technique [0002] Lung cancer has become one of the main causes of human cancer death. The incidence and mortality of lung cancer are the highest among all tumors, but lung cancer is not the most diagnosed tumor. Under the current medical system, the five-year survival rate of patients with advanced lung cancer is only 10-20%, but for the treatment of patients with early-stage lung cancer, about 80-90% of them can be cured by surgical resection. However, the vast majority of early lung cancer patients have no obvious clinical symptoms. When lung cancer is first diagnosed, 85% of patients are in the middle and late stage, only about 14% are diagnosed in the early stage, and only about 16% of th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6806C12Q1/6858C12N15/11
CPCC12Q1/6886C12Q1/6806C12Q1/6858C12Q2600/154C12Q2600/118
Inventor 亓立峰
Owner 上海纳奥生物科技有限公司
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